TGN46 antibody
Sheep anti Human TGN46 antibody (AHP500) used for the identification of the trans golgi network in MD-MB-231 cells by immunofluorescence.
Image caption:
APP redistributes from endosomes to the TGN upon overexpression of μ4-F255A-HA. MD-MB-231 cells were cotransfected with a plasmid encoding either of the indicated HA-epitope-tagged variants of μ4, and with a plasmid encoding APP-GFP carrying the double mutation F615P/D664A. After 24-h cells were fixed, permeabilized, stained for EEA1 and TGN46, and examined by fluorescence microscopy. Merging green, red, and blue channels generated the fourth image on each row; yellow indicates overlapping localization of the green and red channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of the red, green, and blue channels. Insets show 2× magnifications. Bar, 10 μm.
From: Citation: Ross BH, Lin Y, Corales EA, Burgos PV, Mardones GA (2014)
Structural and Functional Characterization of Cargo-Binding Sites on the μ4-Subunit of Adaptor Protein Complex 4.
PLoS ONE 9(2): e88147.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the identification of the trans golgi network in MD-MB-231 cells by immunofluorescence.
Image caption:
APP redistributes from endosomes to the TGN upon overexpression of μ4-R283D-HA or μ4-R283D. MD-MB-231 cells were cotransfected with a plasmid encoding either of the indicated HA-epitope-tagged or untagged variants of μ4, and with a plasmid encoding APP-GFP carrying the double mutation F615P/D664A. After 24-h cells were fixed, permeabilized, stained for EEA1 and TGN46, and examined by fluorescence microscopy. Merging green, red, and blue channels generated the fourth image on each row; yellow indicates overlapping localization of the green and red channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of the red, green, and blue channels. Insets show 2× magnifications. Bar, 10 μm.
From: Citation: Ross BH, Lin Y, Corales EA, Burgos PV, Mardones GA (2014)
Structural and Functional Characterization of Cargo-Binding Sites on the μ4-Subunit of Adaptor Protein Complex 4.
PLoS ONE 9(2): e88147.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the identification of the trans golgi network in H4 Neuroglioma cells by immunofluorescence.
Image caption:
APP redistributes from endosomes to the TGN upon overexpression of μ4-F255A. H4 neuroglioma cells were cotransfected with a plasmid encoding either of the indicated variants of μ4, and with a plasmid encoding APP-GFP carrying the double mutation F615P/D664A. After 36-h cells were fixed, permeabilized, stained for TGN46 and EEA1, and examined by fluorescence microscopy. Merging green, red, and blue channels generated the fourth image on each row; yellow indicates overlapping localization of the green and red channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of the red, green, and blue channels. Insets show 2× magnifications. Bar, 10 μm.
From: Citation: Ross BH, Lin Y, Corales EA, Burgos PV, Mardones GA (2014)
Structural and Functional Characterization of Cargo-Binding Sites on the μ4-Subunit of Adaptor Protein Complex 4.
PLoS ONE 9(2): e88147.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to visualize the cellular distribution of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Cellular distribution of tetherin. (A) Confocal analysis of HeLa cells showing the distribution of endogenous tetherin, detected with a specific antiserum. Cells that were fixed but not permeablized (left panel) allowed visualization of tetherin at the cell surface, while permeabilized cells revealed tetherin concentrated in a perinuclear compartment that was visible in ~50% of cells. This intracellular pool co-localized with a marker for the TGN (TGN-46), as shown by the PDM analysis in the upper right corner of the merged image, where positive co-localization is pseudocolored in orange. Scale bars represent 10 μM. (B) 293A cells were co-transfected with 10 μg HIV-1-pack and 100 ng of expression plasmids for either untagged tetherin or EGFP-tagged tetherin. Cell lysates were analyzed by Western blotting, using antibodies against GFP and tetherin. (C) Cell lysates and pelleted supernatant fractions (VLPs) from same experiment as (B) were probed for HIV-1 p24 expression. Both tetherin constructs inhibited VLP release. (D) HeLa and 293A cells were transfected with either 100 ng or 300 ng of the EGFP-tetherin plasmid. With 300 ng, a punctate pattern of EGFP fluorescence was observed throughout the cells; with 100 ng, the protein could only be detected using an anti-GFP antibody, that revealed an intense surface rim and a fainter PNC in both types of cells. Cells were fixed and permeabilized before staining. Scale bars represent 10 μM. (E) The intracellular concentration of EGFP-tetherin in transiently transfected HeLa cells (100 ng plasmid) was analyzed by confocal microscopy using anti-GFP antibody and specific markers for the TGN (TGN46) and recycling endosomes (endocytosed transferrin). The degree of co-localization was calculated using Pearson's coefficients. Mean +/- SEM is shown for 20 individual cells analyzed.
From: Hauser H, Lopez LA, Yang SJ, Oldenburg JE, Exline CM, Guatelli JC, Cannon PM. HIV-1 Vpu and HIV-2 Env counteract BST-2/tetherin by sequestration in a perinuclear compartment.
Retrovirology. 2010 Jun 7;7:51.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to visualize the cellular distribution of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Redistribution of tetherin to an intracellular compartment by HIV anti-tetherin factors. (A) The percentage of HeLa cells displaying tetherin concentrated in a perinuclear compartment (PNC) was calculated for 100 cells, from either control (Ctrl.) cells or cells transfected with 2 μg of Vpu or ROD10 Env expression plasmids. Mean +/- SEM is shown for n = 2 independent experiments. (B) HeLa cells transfected with either Vpu (Vphu-HcRed) or ROD10 Env, showed increased concentration of tetherin in a perinuclear compartment (arrowed), that co-stained with the TGN marker, TGN46. The triple color merged image is shown. Scale bars represent 10 μM.
From: Hauser H, Lopez LA, Yang SJ, Oldenburg JE, Exline CM, Guatelli JC, Cannon PM. HIV-1 Vpu and HIV-2 Env counteract BST-2/tetherin by sequestration in a perinuclear compartment.
Retrovirology. 2010 Jun 7;7:51.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to visualize the cellular distribution of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Tetherin redistribution by HIV-1 and HIV-2 proviral clones. HeLa cells were either mock treated or transfected with 8 μg of HIV-1NL4-3 or HIV-2ROD10 proviral clones. Cells were fixed, permeabilized, and stained for endogenous tetherin (green), the TGN46 marker (blue), HIV-1 Vpu (red) or HIV-2 Env (red). Triple color merged images are shown. NL4-3 transfected cells showed tetherin co-localized with Vpu and the TGN. ROD10 transfected cells had two distinct appearances. ~25% of cells showed tetherin localized with a compact TGN marker (upper panels), while the majority of the cells had tetherin in a more diffuse perinuclear location that co-localized with more distorted TGN staining (lower panels). Scale bars represent 10 μM.
From: Hauser H, Lopez LA, Yang SJ, Oldenburg JE, Exline CM, Guatelli JC, Cannon PM. HIV-1 Vpu and HIV-2 Env counteract BST-2/tetherin by sequestration in a perinuclear compartment.
Retrovirology. 2010 Jun 7;7:51.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to visualize the cellular distribution of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Analysis of perinuclear concentration of tetherin by Vpu and ROD10 Env. (A) HeLa cells were incubated with anti-tetherin antibody at 4°C for 1 hour, followed by incubation at 37°C for either 15 or 45 minutes to allow endocytosis of antibody-tetherin complexes. The antibody was found in a perinuclear region by 15 minutes (arrowed), but became more diffuse by 45 minutes, suggesting that tetherin quickly exits this compartment and recycles back to the plasma membrane. Scale bars represent 10 μM. (B) HIV-1 VLPs produced in 293A cells in the presence of 100 ng of wild-type (WT) or YY-AA tetherin, with and without co-transfection of 2 μg Vpu (pCMV-Vphu). Expression of proteins was confirmed by Western blotting with specific antibodies; tetherin levels were visualized for both untreated and PNGase F treated cell lysates. Mean +/- SEM fold-enhancement of VLP release by Vpu is shown in presence of tetherin or tetherin (YY-AA), n = 2. (C) HeLa cells expressing EGFP-tetherin or EGFP-tetherin(YY-AA) were incubated for 1.5 hours, with or without 20 μg/ml cycloheximide, and subsequently fixed, permeabilized, and stained with anti-EGFP antibody. In the absence of drug, both WT and YY-AA EGFP-tetherin proteins were found at the cell surface and in a perinuclear region (arrowed). After cycloheximide treatment, only EGFP-tetherin produced this population, with EGFP-tetherin(YY-AA) forming a more punctuate pattern, dispersed throughout the cytoplasm. Two different cells are shown for the drug treatment. Scale bars represent 10 μM. (D) HeLa cells were co-transfected with 60 ng EGFP-tetherin(YY-AA) and 2 μg of expression plasmids for either Vphu-HcRed or ROD10 Env. Cells were incubated with or without cycloheximide, as above, and subsequently fixed, permeabilized, and stained with anti-EGFP (green), anti-TGN46 (blue) and anti-HIV-2 Env (red) antibodies. Vpu was detected by HcRed expression (red). EGFP-tetherin(YY-AA) was concentrated in a perinuclear compartment by Vpu or ROD10 Env, overlapping with the TGN marker, irrespective of cycloheximide treatment. Scale bars represent 10 μM.
From: Hauser H, Lopez LA, Yang SJ, Oldenburg JE, Exline CM, Guatelli JC, Cannon PM. HIV-1 Vpu and HIV-2 Env counteract BST-2/tetherin by sequestration in a perinuclear compartment.
Retrovirology. 2010 Jun 7;7:51.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 is present on RAB6 positive transport carriers. (A) HeLa cells expressing GFP-C11ORF24 were imaged every second for 5 minutes by spinning disk microscopy. The red arrow indicates a small transport carrier whereas the green arrow point to a long tube connected to the Golgi apparatus. A magnified view of these structures is shown in line 2 and 3. (B) HeLa cells were fixed and co-stained with TGN46, C11ORF24 and GTP-RAB6. Colocalization is very strong between TGN46, C11ORF24 and GTP-RAB6. A line profile of the C11ORF24 (green), TGN46 (red) and GTP-RAB6 (blue) fluorescence intensities from the lines in the magnified views are shown in column 7. (C) HeLa cells expressing GFP-C11ORF24 and mCherry-RAB6 were imaged every second for 5 minutes by spinning disk microscopy. The arrow indicates a transport carrier positive for GFP-C11ORF24 (green) and for mCherry-RAB6 (red). A magnified view of this structure is shown lines 4-6. (D) Transport carriers were visualized using a kymograph that was made along the line drawn on the merge image. All the dynamic elements positive for GFP-C11ORF24 (left and merge green) were positive for mCherry-RAB6 (middle and merge red).
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013) C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 localization after Brefeldin A treatment. (A) HeLa cells were either untreated (Control), treated with Brefeldin A for 5 minutes or treated with Brefeldin A for 60 minutes. Cells were then fixed and stained with anti-C11ORF24 (green) and anti-TGN46 (red) antibodies. (B) HeLa cells were treated with cycloheximide and then either directly processed (monensin 0 min) or treated with monensin for 60 minutes. Cells were then fixed and stained with anti-C11ORF24 (green) and anti-GM130 (red) antibodies.
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013) C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-glogi network in COS-1 cells by immunofluorescence.
Image caption:
Co-localization analysis of SMAP2 with several RE/TGN proteins. COS-1 cells were fixed with TCA, permeabilized, and then stained for SMAP2 (green) and the indicated proteins (red). For evection-2, Myc-tagged evection-2 was transiently expressed and stained with anti-Myc antibody. For CTxB, cells were pulsed for 3 min with Alexa 594-CTxB, chased for 35 min at 37°C, and fixed. Fluorescence intensity profile along white dotted lines is shown in the right column. Blue lines in the graph indicate regions where SMAP2 do not co-localize with TfnR or TGN46. Pearson coefficient was obtained using multiple images (n > 12 cells). Data represent mean ± SD. Scale bars, 1 µm.
From: Matsudaira T, Uchida Y, Tanabe K, Kon S, Watanabe T, Taguchi T, et al. (2013) SMAP2 Regulates Retrograde Transport from Recycling Endosomes to the Golgi.
PLoS ONE 8(7): e69145.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in Vero cells by immunofluorescence.
Image caption:
APP relocalizes in infected cells. (A) APP (red) co-localizes with TGN46 (green) in a compact tuft to one side of the nucleus (DAPI, blue) representing the trans-Golgi network in mock infected cells. (B) In cells infected with VP26-GFP HSV1 (green), APP (red) is distributed in particles throughout the cytoplasm at 7 hr p.i. Nuclei are stained with DAPI (blue). (C) Western blotting of uninfected (u) and infected (i) cells with anti-APP, anti-actin (loading control) and anti-VP5 (viral capsid) demonstrates significant increased amound of APP in infected cells. Actin bands remain similar, and VP5, as expected, is only detected in lanes loaded with infected cell lysate. Note no new APP bands are detected in the infected versus uninfected cell lysates. (D) Isolated virus separated on 7.5% SDS-PAGE and stained with amido black for protein and probed for APP by Western blotting with the same antibodies used for immuno-fluorescence. Note that only the 90–110 kD doublet representing APP is detected by anti-APP, with no additional viral protein bands detected. See also Figure S2 for split channels and Figure S3 for histone staining.
From: Cheng S-B, Ferland P, Webster P, Bearer EL (2011) Herpes Simplex Virus Dances with Amyloid Precursor Protein while Exiting the Cell.
PLoS ONE 6(3): e17966.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the immunolocalization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Time course of ShigaB sulfation in annexin A1 or A2 depleted cells. (A) HeLa cells transfected with control or annexin A1 siRNA for 72 h were analyzed for the amount of ShigaB being sulfated at different time points after addition of StxB. Sulfation data are plotted as percentages of the amount of sulphated ShigaB after 30 min incubation with control siRNA treated cells. Black and white bars represent sulfation of ShigaB in control and annexin A1 siRNA treated cells, respectively. Data presented are the average of 3–4 independent experiments, each performed in parallel, error bars indicating standard error of the mean, *p<0.05; **p<0.005 indicates statistically significant change. (B) After the same treatment as in (A), cells were fixed and stained with antibodies against TGN46 and ShigaB. Top panel shows representative confocal pictures for 30 min incubation with StxB, scale bars 20 μm. Left graphic shows quantification of amount of ShigaB colocalized with TGN46. In the right graphic, mean intensity of ShigaB in the Golgi area for the same representative experiment is plotted as percentage of mean intensity in the whole cell. Data presented are the average of at least 35 cells per condition. Quantifications where obtained with ImageJ software, error bars indicating standard error of the mean.
From: Tcatchoff L, Andersson S, Utskarpen A, Klokk TI, Skånland SS, Pust S, et al. (2012)
Annexin A1 and A2: Roles in Retrograde Trafficking of Shiga Toxin.
PLoS ONE 7(7): e40429.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the immunolocalization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Stx transport in annexin A1 depleted cells is regulated by PKCδ and PLA2. (A) Golgi transport of Shiga toxin was evaluated as described in materials and methods by quantification of sulfated ShigaB in HeLa cells transfected with siRNA against annexin A1 or non targeting siRNA, pretreated with the indicated inhibitors. Data from Stx sulfation are plotted as percentages of the value obtained for HeLa cells transfected with control siRNA and treated with DMSO. The white and black bars represent ShigaB-sulf2 sulfation for control and annexin A1 knockdown cells respectively. Data presented are the average of 3 independent experiments, each performed in parallel, error bars indicating standard error of the mean. *p<0.05 indicates statistically significant change between annexin A1 knockdown cells and the corresponding control siRNA treated cells. (B) After treatment with either 5 µM ONO-RS-082 for 30 min or 30 µM MAFP for 1 h, HeLa cells were incubated for 30 min with ShigaB before fixation and staining as indicated in the materials and methods section with antibodies against TGN46 and ShigaB. Panel shows representative confocal pictures, scale bars 20 μm. Graphic shows quantification of amount of ShigaB colocalized with TGN46 in one representative experiment plotted as percentage of control condition. Data presented for one representative experiment (n = 3) are the average of at least 30 cells per condition. Quantifications were obtained with Zen 2009 software from Zeiss, error bars indicating standard error of the mean.
From: Tcatchoff L, Andersson S, Utskarpen A, Klokk TI, Skånland SS, Pust S, et al. (2012)
Annexin A1 and A2: Roles in Retrograde Trafficking of Shiga Toxin.
PLoS ONE 7(7): e40429.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the immunolocalization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Distribution of MPRs and TPST in annexin A1 depleted cells. (A) To visualize the localization of MPRs, cells were fixed, permeabilized and immunostained with a sheep anti-TGN antibody in combination with a mouse monoclonal anti-CD or -CI MPR antibody. In (B), cells were transfected with an EGFP-TPST2 expression plasmid 48 h after transfection with siRNA. TGN was stained as in (A). Scale bars 10 μm. The relative colocalization of MPR46, MPR300 or EGFP-TPST2 with TGN46 positive structures was quantified by ImageJ software. Graphs represent the average from 25 cells plotted as percentage of total fluorescence for each marker, for one representative experiment (n = 3), where error bars indicate standard deviation.
From: Tcatchoff L, Andersson S, Utskarpen A, Klokk TI, Skånland SS, Pust S, et al. (2012)
Annexin A1 and A2: Roles in Retrograde Trafficking of Shiga Toxin.
PLoS ONE 7(7): e40429.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
The expanded Golgi remains polarized. HeLa cells were transfected with control (ctrl) or Scyl1 inhibitory miRNAs that were driven from a plasmid that also expresses emGFP. The cells were subsequently fixed and processed for indirect immunofluorescence with antibodies against TGN46 (red) and GM130 (blue). The box in the left panels indicates the expanded area shown in the right panels. The scale bar = 10 μm for the left and 4 μm for the right images, respectively.
From: Burman JL, Hamlin JNR, McPherson PS (2010)
Scyl1 Regulates Golgi Morphology.
PLoS ONE 5(3): e9537.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in M10 cells by immunofluorescence
Image caption:
Over-expression of LAPTM5 does not affect the cellular distribution of CD1e molecules. A) Fixed, permeabilized M10 cells expressing CD1e alone or co-expressing CD1e and EYFP-LAPTM5 were stained with the anti-CD1e mAb 20.6 and antibodies specific for TGN46, EEA1, CD63 or HLA-DR. B) Transfected HEK293 cells expressing CD1e alone or co-expressing LAPTM5-V5 were fixed, permeabilized and stained with the anti-CD1e mAb 20.6 and antibodies specific for TGN46, EEA1 and CD63. Scale bar, 10 μM.
From: Angénieux C, Waharte F, Gidon A, Signorino-Gelo F, Wurtz V, Hojeij R, et al. (2012)
Lysosomal-Associated Transmembrane Protein 5 (LAPTM5) Is a Molecular Partner of CD1e.
PLoS ONE 7(8): e42634.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in human cell lines by immunofluorescence.
Image caption:
Vpu ELV mutants localize to early endosomal compartments in tetherin-expressing cells. 293T/tetherin (A), HeLa (B), Jurkat (C) or parental 293T (D) were infected with HIV-1 wt or HIV-1 Vpu ELV at an MOI of 1. 48 h later the cells were fixed and stained for Vpu (green) and the TGN marker TGN46 (red) and examined by confocal microscopy. Panels are of representative examples. (E) The percentage of the total Vpu immunoreactivity localized to TGN46+ compartments was calculated for cells (n = 20) from A–D using the Leica Confocal Software. Results were analyzed by unpaired 2-tailed t-test - ** P = 10−8 or lower. (F) HeLa cells as in B were stained for Vpu (green) and the early endosomal marker EEA1 or late endosomal marker CD63 (red).
From: Kueck T & Neil SJD (2012)
A Cytoplasmic Tail Determinant in HIV-1 Vpu Mediates Targeting of Tetherin for Endosomal Degradation and Counteracts Interferon-Induced Restriction.
PLoS Pathog 8(3): e1002609.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in human cell lines by immunofluorescence.
Image caption:
The second alpha helix of Vpu can be functionally replaced by the D/EXXXLL endocytic signal from HIV-1 Nef. (A) Schematic representation of Vpu ENTSLL and Vpu-Nef chimeric constructs. (B) 293T cells were transfected with NL4.3 wt of NL4.3 delVpu proviral plasmids in combination with tetherin and indicated pCR3.1 Vpu-HA or Vpu-Nef-HA chimera. 48 h post transfection, cell lysates and pelleted supernatant virions were harvested and subjected to SDS-PAGE and analyzed by Western blotting for HIV-1 p24CA, Vpu-HA and Hsp90, and analyzed by LiCor quantitative imager. (C) Infectivity of viral supernatants from B was determined on HeLa-TZMbl cells as in Figure 1B. Error bars represent standard deviation of three independent experiments. (D) HeLa cells were co-transfected with Vpu or indicated Vpu mutant and a GFP expression construct. Cell surface staining for endogenous tetherin was analyzed by flow cytometry 48 h post transfection, as in Figure 2D. (E) HeLa cells were transfected with Vpu-Nef or Vpu-Nef-mu CherryFP fusions (red) and counterstained for TGN46 (green) and DAPI (blue) and examined by confocal microscopy.
From: Kueck T & Neil SJD (2012)
A Cytoplasmic Tail Determinant in HIV-1 Vpu Mediates Targeting of Tetherin for Endosomal Degradation and Counteracts Interferon-Induced Restriction.
PLoS Pathog 8(3): e1002609.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in human cell lines by immunofluorescence.
Image caption:
Effects of Vpu on surface expression of human tetherin in murine fibroblasts deficient for AP3 or AP1. Fibroblasts from pearl (AP3δ−/−) (A) or AP-1μ1A−/− mice (B) or their reconstituted counterparts were transduced to express human tetherin bearing an extracellular HA-tag. The cells were then transduced with retroviral vector constructs encoding Vpu linked to GFP via an IRES. 48 h later the cells were surface stained for human tetherin expression using anti-HA antibodies. (C) HeLa-CD8-CI-M6PR cells were treated with control or Vps26 siRNAs. 48 h later the cells were fixed and stained for CD8 (green) and TGN46 (red).
From: Kueck T & Neil SJD (2012)
A Cytoplasmic Tail Determinant in HIV-1 Vpu Mediates Targeting of Tetherin for Endosomal Degradation and Counteracts Interferon-Induced Restriction. PLoS Pathog 8(3): e1002609.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in melanoma cells by immunofluorescence.
Image caption:
Amino acids 834-841 of the gHcyt are dispensable for intracellular trafficking and endocytosis of gH in vitro. Confocal microscopy of melanoma cells transiently expressing gH[WT] or gH mutants with gL at 24 hours post transfection. Cells were stained for gH (red), early endosome antigen (EEA1; green), trans-Golgi network (TGN46; green), and nuclei (Hoechst 33342; blue). White arrows indicate colocalization of EEA1 and gH, which highlight representative endocytic vesicles containing gH. The scale bars represent 20 μm.
From: Yang E, Arvin AM & Oliver SL (2014)
The Cytoplasmic Domain of Varicella-Zoster Virus Glycoprotein H Regulates Syncytia Formation and Skin Pathogenesis.
PLoS Pathog 10(5): e1004173.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to identification of the trans-Golgi network in wild type and &alpha-synuclein overexpressing mouse neurons.
Image caption:
MPR300 preferentially localizes in late endosomes in ASOTg/Tg neurons.
(A-C) Confocal microscopy analysis of MPR300 (green) and EEA1, TGN46 and Rab7 (red), respectively, in WT and ASOTg/Tg cortical neurons. Scale bar 5μm. (D) The extent of MPR300 co-localization to EEA1, TGN46 and Rab7 is reported in panel D. Quantitative analysis was performed using Zen software. The (R) coefficient (Pearson's coefficient) was used for the quantitative and comparative analyses (R). Data are expressed as meaN ± SEM. n = 8. *p<0.05. Two tailed T-Test).
From: Matrone C, Dzamko N, Madsen P, Nyegaard M, Pohlmann R, Søndergaard RV, et al. (2016)
Mannose 6-Phosphate Receptor Is Reduced in -Synuclein Overexpressing Models of Parkinsons Disease.
PLoS ONE 11(8): e0160501.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500) used to visualize the trans golgi network in HeLa cells by immunofluorescence.
Image caption:
Membrane-proximal, hinge region E28/L33 residues are important for Vpu localization in the TGN. Intracellular localization of the Vpu mutants. HeLa cells were transfected with proviral plasmids encoding either WT Vpu or one of E28A/L33A, E59K/L63F, E28A/L33A-E59K/L63F and A10L/A14L/A18L Vpu mutants and were co-stained with anti-TGN46 (red, for TGN) and anti-Vpu (green) Abs as well as with DAPI (grey, for nucleus). Shown are representative confocal microscopy pictures for each of the Vpu mutants, with two prototypical patterns (1&2) of localization observed with the E28A/L33A mutant. The white bars represent a distance of 10 μm.
From: Lukhele S & Cohen ÉA.
Conserved residues within the HIV-1 Vpu transmembrane-proximal hinge region modulate BST2 binding and antagonism.
Retrovirology. 2017 14(1):18.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500) used to visualize the trans golgi network in HeLa cells by immunofluorescence.
Image caption:
E28/L33 residues have no intrinsic activity on Vpu cellular distribution in the absence of BST2. a Intracellular localization of Vpu mutants in HeLa cells depleted of BST2 (SH-BST2, treated with shRNA against BST2) or not (NT-BST2, treated with non-targeting shRNA). Transfected cells were co-stained with anti-TGN46 (red, for TGN) and anti-Vpu (green) Abs as well as with DAPI (grey, for nucleus). Shown are representative confocal microscopy pictures for each of the Vpu mutants. The white bars represent a distance of 10 μm.
From: Lukhele S & Cohen ÉA.
Conserved residues within the HIV-1 Vpu transmembrane-proximal hinge region modulate BST2 binding and antagonism.
Retrovirology. 2017 14(1):18.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500) used to visualize the trans golgi network in HeLa cells by immunofluorescence.
Image caption:
Hinge region E28/L33 residues are important for Vpu co-localization with BST2. a Representative pictures showing extent of co-localization of Vpu mutants with endogenous BST2 in HeLa cells as determined by the co-staining of anti-Vpu (green) and anti-BST2 (red) Abs following intracellular staining for both Vpu and BST2. The white bars in panel A represent a distance of 10 μm
From: Lukhele S & Cohen ÉA.
Conserved residues within the HIV-1 Vpu transmembrane-proximal hinge region modulate BST2 binding and antagonism.
Retrovirology. 2017 14(1):18.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500) used for the demonstration of the Golgi apparatus in MDA-MB-231 cells by immunofluorescence.
Image caption:
Effect of Golgi disrupting treatments on the Golgi apparatus of MDA-MB-231 cells.
Cells were left untreated (A; Control), or transfected to transiently express the HA-epitope-tagged ARF1 constitutively-activated mutant for 16 h (B; HA-ARF1-Q71L), or treated for 60 min either with 5 μg/ml Brefeldin A (C; BFA) or 10μM Golgicide A (D; GCA). Cells were fixed, permeabilized, and immunolabeled with mouse monoclonal antibody to GM130, rabbit polyclonal antibody to Giantin, and sheep antibody to TGN46. Secondary antibodies were Alexa-594-conjugated donkey anti-mouse IgG (red channel), Alexa-488-conjugated donkey anti-rabbit IgG (green channel), and Alexa-647-conjugated donkey anti-sheep IgG (blue channel). Nuclei were stained with DAPI (gray channel). Stained cells were examined by fluorescence microscopy. Merging red, green, blue, and grey channels generated the fourth image on each row; yellow indicates overlapping localization of the red and green channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of all three channels. Bar, 10μm.
From: Citation: Luchsinger C, Aguilar M, Burgos PV, Ehrenfeld P, Mardones GA (2018)
Functional disruption of the Golgi apparatus protein ARF1 sensitizes MDA-MB-231 breast cancer cells to the antitumor drugs Actinomycin D and Vinblastine through ERK and AKT signaling.
PLoS ONE 13(4): e0195401.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500) used for the demonstration of the Golgi apparatus in MDA-MB-231 cells by immunofluorescence.
Image caption:
Effect of Actinomycin D and Vinblastine on the Golgi apparatus of MDA-MB-231 cells.
Cells were left untreated (A; Control), or treated for 60 min either with 10 ng/ml Actinomycin D (B; ActD) or 25 nM Vinblastine (C; VLB). Cells were fixed, permeabilized, and immunolabeled with mouse monoclonal antibody to GM130, rabbit polyclonal antibody to Giantin, and sheep antibody to TGN46. Secondary antibodies were Alexa-594-conjugated donkey anti-mouse IgG (red channel), Alexa-488-conjugated donkey anti-rabbit IgG (green channel), and Alexa-647-conjugated donkey anti-sheep IgG (blue channel). Nuclei were stained with DAPI (gray channel). Stained cells were examined by fluorescence microscopy. Merging red, green, blue, and grey channels generated the fourth image on each row; yellow indicates overlapping localization of the red and green channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of all three channels. Bar, 10μm.
From: Citation: Luchsinger C, Aguilar M, Burgos PV, Ehrenfeld P, Mardones GA (2018)
Functional disruption of the Golgi apparatus protein ARF1 sensitizes MDA-MB-231 breast cancer cells to the antitumor drugs Actinomycin D and Vinblastine through ERK and AKT signaling.
PLoS ONE 13(4): e0195401.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to identigy the Golgi apparatus in human skin fibroblasts in vitro by immunofluorescence.
Image caption:
(A) Skin fibroblasts of subject 3 (S3) carrying the homozygous p.Gly296Arg variant show intracellular ADAMTSL2 mislocalization. Representative confocal immunofluorescence images of permeabilized fibroblasts stained for ADAMTSL2 (green) and TGN46 (red), a trans-Golgi cisternae marker. In wild-type (WT) cells, ADAMTSL2 has diffuse cytoplasmatic signal, with a strong positive signal at the Golgi level where co-localization with TGN46 occurs, whereas fibroblasts from S3 display a focal cytoplasmatic distribution of ADAMTSL2 which fails to co-localize with TGN46. Nuclei were counterstained with DAPI (blue).
From: Piccolo P, Sabatino V, Mithbaokar P, Polishchuck E, Law SK, Magraner-Pardo L, Pons T, Polishchuck R, Brunetti-Pierri N.
Geleophysic dysplasia: novel missense variants and insights into ADAMTSL2 intracellular trafficking.
Mol Genet Metab Rep. 2019 Sep 5;21:100504.
doi:10.1016/j.ymgmr.2019.100504.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in M10 cells by immunofluorescence
Image caption:
Cellular distribution of LAPTM5. Transfected M10 cells co-expressing EYFP-LAPTM5 and CD1e were fixed, permeabilized and co-stained with the anti-CD1e mAb 20.6, revealed using Cy3-conjugated antibodies (A) and antibodies specific for TGN46, revealed by Cy5-conjugated antibodies (B, left panel, represented using red pseudo-color), or stained with anti-EEA1, -CD63, -Lamp1 (B) or -HLA-DR Abs (C). Untreated cells (upper panel) were compared with cells treated for 4 hours with bafilomycin (lower panel) (A). D) M10 cells co-expressing CD1e and EYFP-LAPTM5 were fixed and processed for immunolabeling of cryosections with 10 nm anti-GFP and 15 nm anti-CD1e conjugated gold particles. EYFP-LAPTM5 and CD1e were detected in the endoplasmic reticulum (ER), in Golgi compartments (Golgi) and in multivesicular (MVs) and multilamellar endosomes (MLs). E, F) Characterization of two anti-LAPTM5 mAbs. M10 cells co-expressing CD1e and EYFP-LAPTM5 were fixed, permeabilized and stained with an anti-LAPTM5 mAb (2.16.1 or 47.15.6), which was revealed with Cy3-conjugated secondary Abs (E). M10 cells expressing CD1e were fixed, permeabilized and double-labeled with 2.16.1 and either anti-TGN46, anti-EEA1, anti-CD63 (H5C6) or anti-HLA-DR (L243) Abs (F). Scale bars, confocal micrographs, 10 μM; electron microscopy micrographs, 100 nM.
From: Angénieux C, Waharte F, Gidon A, Signorino-Gelo F, Wurtz V, Hojeij R, et al. (2012)
Lysosomal-Associated Transmembrane Protein 5 (LAPTM5) Is a Molecular Partner of CD1e.
PLoS ONE 7(8): e42634.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Disruption of the golgin network displaces Scyl1 from the Golgi. A, HeLa cells were transfected with control or p115 siRNA and were subsequently fixed and processed for indirect immunofluorescence with antibodies against Scyl1 (red) and GM130 (green). The area indicated by the box in the merged image is shown at higher power on the right. Arrowheads indicate co-localizing structures. The scale bar = 10 μm for the low and 1 μm for high power images, respectively. B, HeLa cells were transfected with control or p115 siRNA and with ERGIC53-YFP, and were subsequently fixed and processed for indirect immunofluorescence with antibodies against Scyl1 (red) and GM130 (blue) with ERGIC53 imaged in the green channel; top eight panels or with antibodies against TGN46 (red), GM130 (green) and Scyl1 (blue); bottom 4 panels. For the middle four panels, arrowheads indicate co-localizing structures. For the bottom four panels, arrowheads indicate areas where TGN46 and GM130 are found in close apposition and lack the presence of Scyl1. The scale bar = 1 μm.
From: Burman JL, Hamlin JNR, McPherson PS (2010)
Scyl1 Regulates Golgi Morphology.
PLoS ONE 5(3): e9537.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 is localized on the trans-Golgi Network. (A) HeLa cells were transfected with a construct encoding human C11ORF24 tagged with GFP and fixed 18 hours later. Cells were co-stained with TGN46 (middle, red) and GM130 (right, blue). The second line represents a closer view of the Golgi apparatus outlined by a rectangle in the merge image. (B) HeLa cells were fixed and co-stained with TGN46, C11ORF24 and Giantin. Colocalization is very strong between TGN46 and C11ORF24 (column 4) whereas it is lower with Giantin (column 5). Line profiles of the C11ORF24 (green), TGN46 (red) and Giantin (blue) fluorescence intensities from the lines in the magnified views are shown in column 7. The colocalization between TGN46 and C11ORF24 is clearly visible throughout mitosis (lines 2-5). (C) HeLa cells were stained for C11ORF24 and imaged using dSTORM super-resolution microscopy. A Fire LUT was used to facilitate visualization. A magnified view of the boxed region of the left image is shown in the middle and a magnified view of the boxed region of the middle image is shown on the right. The cell contour is outlined in red.
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013)
C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the identification of the trans golgi network in MD-MB-231 cells by immunofluorescence.
Image caption:
APP redistributes from endosomes to the TGN upon overexpression of μ4-R283D-HA or μ4-R283D. MD-MB-231 cells were cotransfected with a plasmid encoding either of the indicated HA-epitope-tagged or untagged variants of μ4, and with a plasmid encoding APP-GFP carrying the double mutation F615P/D664A. After 24-h cells were fixed, permeabilized, stained for EEA1 and TGN46, and examined by fluorescence microscopy. Merging green, red, and blue channels generated the fourth image on each row; yellow indicates overlapping localization of the green and red channels, cyan indicates overlapping localization of the green and blue channels, magenta indicates overlapping localization of the red and blue channels, and white indicates overlapping localization of the red, green, and blue channels. Insets show 2× magnifications. Bar, 10 μm.
From: Citation: Ross BH, Lin Y, Corales EA, Burgos PV, Mardones GA (2014)
Structural and Functional Characterization of Cargo-Binding Sites on the μ4-Subunit of Adaptor Protein Complex 4.
PLoS ONE 9(2): e88147.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the immunolocalization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Distribution of MPRs and TPST in annexin A1 depleted cells. (A) To visualize the localization of MPRs, cells were fixed, permeabilized and immunostained with a sheep anti-TGN antibody in combination with a mouse monoclonal anti-CD or -CI MPR antibody. In (B), cells were transfected with an EGFP-TPST2 expression plasmid 48 h after transfection with siRNA. TGN was stained as in (A). Scale bars 10 μm. The relative colocalization of MPR46, MPR300 or EGFP-TPST2 with TGN46 positive structures was quantified by ImageJ software. Graphs represent the average from 25 cells plotted as percentage of total fluorescence for each marker, for one representative experiment (n = 3), where error bars indicate standard deviation.
From: Tcatchoff L, Andersson S, Utskarpen A, Klokk TI, Skånland SS, Pust S, et al. (2012)
Annexin A1 and A2: Roles in Retrograde Trafficking of Shiga Toxin.
PLoS ONE 7(7): e40429.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
The expanded Golgi remains polarized. HeLa cells were transfected with control (ctrl) or Scyl1 inhibitory miRNAs that were driven from a plasmid that also expresses emGFP. The cells were subsequently fixed and processed for indirect immunofluorescence with antibodies against TGN46 (red) and GM130 (blue). The box in the left panels indicates the expanded area shown in the right panels. The scale bar = 10 μm for the left and 4 μm for the right images, respectively.
From: Burman JL, Hamlin JNR, McPherson PS (2010)
Scyl1 Regulates Golgi Morphology.
PLoS ONE 5(3): e9537.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 localization after Brefeldin A treatment. (A) HeLa cells were either untreated (Control), treated with Brefeldin A for 5 minutes or treated with Brefeldin A for 60 minutes. Cells were then fixed and stained with anti-C11ORF24 (green) and anti-TGN46 (red) antibodies. (B) HeLa cells were treated with cycloheximide and then either directly processed (monensin 0 min) or treated with monensin for 60 minutes. Cells were then fixed and stained with anti-C11ORF24 (green) and anti-GM130 (red) antibodies.
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013) C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label golgi network in human H4 neuroglioma cells transfected with wildtype RAB1A by immunofluorescence.
Image caption:
Cells were triple-labeled with rabbit polyclonal antibody to GOLPH3, mouse monoclonal antibody to Giantin and sheep polyclonal antibody to TGN46. Secondary antibodies were Alexa-Fluor-594-conjugated donkey anti-rabbit IgG, Alexa-Fluor-647-conjugated donkey anti-mouse IgG and Alexa-Fluor-350-conjugated donkey anti-sheep IgG. Stained cells were examined by fluorescence microscopy. Insets in B: X3 magnification, with arrows indicating colocalization at Golgi punctae. Bar, 10 μm. For comparison of the fluorescence signals, pairs of images were pseudocolored as indicated.
From: Cavieres VA, Cerda-Troncoso C, Rivera-Dictter A, Castro RI, Luchsinger C, Santibañez N, et al. (2020)
Human Golgi phosphoprotein 3 is an effector of RAB1A and RAB1B.
PLoS ONE 15(8): e0237514.
doi:10.1371/journal.pone.0237514
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label golgi network in human H4 neuroglioma cells transfected with GFP-RAB1A-S25N by immunofluorescence.
Image caption:
Cells were triple-labeled with rabbit polyclonal antibody to GOLPH3, mouse monoclonal antibody to Giantin and sheep polyclonal antibody to TGN46. Secondary antibodies were Alexa-Fluor-594-conjugated donkey anti-rabbit IgG, Alexa-Fluor-647-conjugated donkey anti-mouse IgG and Alexa-Fluor-350-conjugated donkey anti-sheep IgG. Stained cells were examined by fluorescence microscopy. Insets in B: X3 magnification, with arrows indicating colocalization at Golgi punctae. Bar, 10 μm. For comparison of the fluorescence signals, pairs of images were pseudocolored as indicated.
From: Cavieres VA, Cerda-Troncoso C, Rivera-Dictter A, Castro RI, Luchsinger C, Santibañez N, et al. (2020)
Human Golgi phosphoprotein 3 is an effector of RAB1A and RAB1B.
PLoS ONE 15(8): e0237514.
doi:10.1371/journal.pone.0237514
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label golgi network in human H4 neuroglioma cells transfected with GFP-RAB1A-Q70L by immunofluorescence.
Image caption:
Cells were triple-labeled with rabbit polyclonal antibody to GOLPH3, mouse monoclonal antibody to Giantin and sheep polyclonal antibody to TGN46. Secondary antibodies were Alexa-Fluor-594-conjugated donkey anti-rabbit IgG, Alexa-Fluor-647-conjugated donkey anti-mouse IgG and Alexa-Fluor-350-conjugated donkey anti-sheep IgG. Stained cells were examined by fluorescence microscopy. Insets in B: X3 magnification, with arrows indicating colocalization at Golgi punctae. Bar, 10 μm. For comparison of the fluorescence signals, pairs of images were pseudocolored as indicated.
From: Cavieres VA, Cerda-Troncoso C, Rivera-Dictter A, Castro RI, Luchsinger C, Santibañez N, et al. (2020)
Human Golgi phosphoprotein 3 is an effector of RAB1A and RAB1B.
PLoS ONE 15(8): e0237514.
doi:10.1371/journal.pone.0237514
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 is localized on the trans-Golgi Network. (A) HeLa cells were transfected with a construct encoding human C11ORF24 tagged with GFP and fixed 18 hours later. Cells were co-stained with TGN46 (middle, red) and GM130 (right, blue). The second line represents a closer view of the Golgi apparatus outlined by a rectangle in the merge image.
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013)
C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to localize the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
C11ORF24 is localized on the trans-Golgi Network. (B) HeLa cells were fixed and co-stained with TGN46, C11ORF24 and Giantin. Colocalization is very strong between TGN46 and C11ORF24 (column 4) whereas it is lower with Giantin (column 5). Line profiles of the C11ORF24 (green), TGN46 (red) and Giantin (blue) fluorescence intensities from the lines in the magnified views are shown in column 7. The colocalization between TGN46 and C11ORF24 is clearly visible throughout mitosis (lines 2-5).
From: Fraisier V, Kasri A, Miserey-Lenkei S, Sibarita J-B, Nair D, Mayeux A, et al. (2013)
C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles.
PLoS ONE 8(12): e82223.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
Disruption of the golgin network displaces Scyl1 from the Golgi. B, HeLa cells were transfected with control or p115 siRNA and with ERGIC53-YFP, and were subsequently fixed and processed for indirect immunofluorescence with antibodies against Scyl1 (red) and GM130 (blue) with ERGIC53 imaged in the green channel; top eight panels or with antibodies against TGN46 (red), GM130 (green) and Scyl1 (blue); bottom 4 panels. For the middle four panels, arrowheads indicate co-localizing structures. For the bottom four panels, arrowheads indicate areas where TGN46 and GM130 are found in close apposition and lack the presence of Scyl1. The scale bar = 1 μm.
From: Burman JL, Hamlin JNR, McPherson PS (2010)
Scyl1 Regulates Golgi Morphology.
PLoS ONE 5(3): e9537.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in M10 cells by immunofluorescence
Image caption:
Over-expression of LAPTM5 does not affect the cellular distribution of CD1e molecules. A) Fixed, permeabilized M10 cells expressing CD1e alone or co-expressing CD1e and EYFP-LAPTM5 were stained with the anti-CD1e mAb 20.6 and antibodies specific for TGN46, EEA1, CD63 or HLA-DR. B) Transfected HEK293 cells expressing CD1e alone or co-expressing LAPTM5-V5 were fixed, permeabilized and stained with the anti-CD1e mAb 20.6 and antibodies specific for TGN46, EEA1 and CD63. Scale bar, 10 μM.
From: Angénieux C, Waharte F, Gidon A, Signorino-Gelo F, Wurtz V, Hojeij R, et al. (2012)
Lysosomal-Associated Transmembrane Protein 5 (LAPTM5) Is a Molecular Partner of CD1e.
PLoS ONE 7(8): e42634.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in melanoma cells by immunofluorescence.
Image caption:
Pathogenesis of VZV with mutations in the gHcyt in human skin xenografts. Confocal micrographs of 5 μm sections of human skin xenografts with the highest titers infected with either pOka or pOka-gH[Δ834-841], -gH[Y835A], -gH[Y835F], -gH[TL], and -gH[V5] at 21 dpi. The sections were stained for capsids (ORF23; green), gE (red), TGN46 (violet), and nuclei (Hoechst 33342; blue). The scale bar for skin implants infected with pOka (top left) and gH[Δ834-841] (top right) represent 200 μm. White squares are shown in higher magnification immediately below (A = pOka, B = Δ834-841). Higher magnification images consist of a four-color merge (top), two color-ORF23 and gE merge (bottom left), and two color-TGN46 and nuclei merge (bottom right). Lesions were not detected for pOka-gH[TL]-infected xenografts. The scale bars for higher magnification images represent 50 μm.
From: Yang E, Arvin AM & Oliver SL (2014)
The Cytoplasmic Domain of Varicella-Zoster Virus Glycoprotein H Regulates Syncytia Formation and Skin Pathogenesis.
PLoS Pathog 10(5): e1004173.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used for the localization of the trans-golgi network in melanoma cells by immunofluorescence.
Image caption:
Amino acids 834-841 of the gHcyt are dispensable for intracellular trafficking and endocytosis of gH during infection of melanoma cells. Confocal microscopy of melanoma cells at 24(Uninfected) or infection with pOka-gH[Δ834-841], pOka-gH[TL], pOka-gH[Y835A], pOka-gH[Y835F], pOka-gH[V5], pOka-gH[834StopV5], or pOka viruses. Cells were stained for gH (red), early endosome antigen (EEA1; green), trans-Golgi network (TGN46; green) and nuclei (Hoechst 33342; blue). White arrows indicate colocalization of EEA1 and gH, which highlight representative endocytic vesicles containing gH. The scale bars represent 20 μm.
From: Yang E, Arvin AM & Oliver SL (2014)
The Cytoplasmic Domain of Varicella-Zoster Virus Glycoprotein H Regulates Syncytia Formation and Skin Pathogenesis.
PLoS Pathog 10(5): e1004173.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to demonstrate the trans golgi network in Vero cells by immunofluorescence.
Image caption:
Perturbation of TGN46 distribution and Golgi morphological changes in ΔSYS1 cells. (B) Effect of SYS1 KO on localization of TGN46. Parent cells, ΔSYS1 cells, and ΔSYS1/SYS-1-HA cells were stained with mouse anti-GM130 Abs and sheep anti-TGN46 (TGN marker) Abs, followed by Alexa Fluor 488-conjugated anti-mouse Abs (green) and Alexa Fluor 594-conjugated anti-sheep Abs (magenta). Scale bars, 10 μm and 2 μm (zoom).
From: Sakuma C, Sekizuka T, Kuroda M, Hanada K, Yamaji T.
Identification of SYS1 as a Host Factor Required for Shiga Toxin-Mediated Cytotoxicity in Vero Cells. Int J Mol Sci. 2021; 22(9):4936.
doi: 10.3390/ijms22094936
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to highlight the trans-Golgi network in Human HEK293 cells by immunofluorescence.
Image caption:
The localization of ATG9 at the late endosome and the trans-Golgi network is reduced with DNAJC13 knockdown.
Confocal images of HEK293A cells transfected with nonsense or DNAJC13 siRNA under steady-state conditions. Cells were stained for ATG9A and (A) trans-Golgi network protein 2 (TGN46).
From: Besemer AS, Maus J, Ax MDA, Stein A, Vo S, Freese C, Nalbach K, von Hilchen C, Pfalzgraf IF, Koziollek-Drechsler I, Silva B, Huesmann H, Boukhallouk F, Florin L, Kern A, Behl C, Clement AM.
Receptor-mediated endocytosis 8 (RME-8)/DNAJC13 is a novel positive modulator of autophagy and stabilizes cellular protein homeostasis.
Cell Mol Life Sci. 2021 Jan;78(2):645-660.
doi: 10.1007/s00018-020-03521-y.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label Golgi apparatus in HeLa cells by immunofluorescence. Detailed in enlargement on right.
Image caption:
Arp2/3 inhibition induces an accumulation of GM130+ and RAB1+ membrane tubules.
HeLa cells co-labeled with antibody against GM130 and antibodies against TGN. Maximum projections of confocal slices around the Golgi are shown. Scale bar: 10μm.
From: Yoshimura, A.; Miserey-Lenkei, S.; Coudrier, E.; Goud, B.
Branched Actin Maintains Acetylated Microtubule Network in the Early Secretory Pathway.
Cells 2022. 11 (1): 15.
doi: 10.3390/cells11010015
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to detect TGN expression in vitro by immunofluorescence.
Image caption:
Western blot analysis of whole cell lysates and lysosome fractions isolated from NHT and USP32 KO cells.
From: Hertel A, Alves LM, Dutz H, Tascher G, Bonn F, Kaulich M, Dikic I, Eimer S, Steinberg F, Bremm A.
USP32-regulated LAMTOR1 ubiquitination impacts mTORC1 activation and autophagy induction.
Cell Rep. 2022 Dec 6;41(10):111653.
doi: 10.1016/j.celrep.2022.111653.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500GT) used to identify the trans-Golgi network by immunofluorescence.
Image caption:
Impact of the Y8A mutation of ATG9A on its sorting and segregation from synaptophysin.
COS7 cells expressing ATG9A-EGFP (WT) or ATG9AY8A-EGFP and immunostained for cis-Golgi (GM130) and trans-Golgi (TGN46) markers, showing that ATG9AY8A is retained in the TGN area, where it colocalizes with TGN46.
From: Park D, Wu Y, Wang X, Gowrishankar S, Baublis A, De Camilli P.
Synaptic vesicle proteins and ATG9A self-organize in distinct vesicle phases within synapsin condensates.
Nat Commun. 2023 Jan 28;14(1):455.
doi: 10.1038/s41467-023-36081-3.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
AP-1 regulates the TGN-to-cell surface delivery of EGFR. B, wt HeLa cells or AP1γ1 KO HeLa cells were transiently transfected with plasmids encoding the indicated constructs. Day 1 after transfection, the cells were incubated with biotin for 2 h, and the localizations of the indicated proteins were analyzed by immunofluorescence . Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y.
The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
AP-1 interacts with Y998RAL motif of EGFR to regulate TGN export of EGFR.
HeLa cells were transiently transfected with SBP-EGFP-EGFR or SBP-EGFP-EGFRY998A. Day 1 after transfection, SBP-EGFP-EGFR or SBP-EGFP-EGFRY998A was released by adding biotin for 2 h. The localizations of the indicated proteins were analyzed by immunofluorescence. Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y.
The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
Rab12 interacts with EGFR and regulates the TGN-to-cell surface delivery of EGFR.
HeLa cells were mock transfected or transfected with siRNA against Rab12 and re-transfected after 24 h with plasmid encoding the indicated constructs. After an additional 24 h, cells were incubated with biotin for 2 h, and the localizations of the indicated proteins were analyzed by immunofluorescence. Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y.
The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
Rab12 interacts with EGFR and regulates the TGN-to-cell surface delivery of EGFR.
HeLa cells were mock transfected or transfected with siRNA against the indicated Rab proteins and re-transfected after 24 h with plasmid encoding SBP-EGFP-EGFR. After an additional 24 h, the localizations of the indicated proteins were analyzed by immunofluorescence. Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y.
The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
Rab12 interacts with EGFR and regulates the TGN-to-cell surface delivery of EGFR.
HeLa cells were mock transfected or transfected with siRNA against the indicated Rab proteins and re-transfected after 24 h with plasmid encoding SBP-EGFP-EGFR. After an additional 24 h, the localizations of the indicated proteins were analyzed by immunofluorescence. Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y.
The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
Surface delivery of EGFRL858Rindependent of AP-1 and Rab12. The localizations of the indicated proteins were analyzed by immunofluorescence in the indicated cells 2 h after biotin treatment. Size bar, 10 μm
Wang J, Lau PK, Li CW, Guo Y. The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
Surface delivery of EGFRL858Rindependent of AP-1 and Rab12. The localizations of the indicated proteins were analyzed by immunofluorescence in the indicated cells 2 h after biotin treatment. Size bar, 10 μm.
From: Wang J, Lau PK, Li CW, Guo Y. The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti TGN46 antibody (AHP500G) used to label the trans-Golgi network by immunofluorescence.
Image caption:
N-terminal and C-terminal HA-tagged Rab12 are located at the TGN. HeLa cells expressing Rab12-HA or HA-Rab12 were incubated with or without 40 μg/mL digitonin for 5 min on ice and washed with PBS. The localizations of the indicated proteins were analyzed by immunofluorescence. Size bar, 10 μm.
Wang J, Lau PK, Li CW, Guo Y. The clathrin adaptor complex-1 and Rab12 regulate post-Golgi trafficking of wild-type epidermal growth factor receptor (EGFR).
J Biol Chem. 2023 Feb 3:102979.
doi: 10.1016/j.jbc.2023.102979.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to label the trans golgi network in human endothlial cells by immunofluorescence.
Image caption:
(H) GBF1 depletion in endothelial cells does not induce gross morphological Golgi defects but causes dispersion of the ERGIC. Immunofluorescence confocal images of control and GBF1-siRNA-treated HUVECs for various ER-Golgi proteins. Scale bars, 10 μm.
From: Lopes-da-Silva M, McCormack JJ, Burden JJ, Harrison-Lavoie KJ, Ferraro F, Cutler DF.
A GBF1-Dependent Mechanism for Environmentally Responsive Regulation of ER-Golgi Transport.
Dev Cell. 2019 Jun 3;49(5):786-801.e6.
doi: 10.1016/j.devcel.2019.04.006.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to label the trans golgi network in human endothlial cells by immunofluorescence.
Image caption:
(I) Super-resolution structured illumination microscopy (SR-SIM) reconstruction of control and GBF1-siRNA-treated cells stained for TGN46, green and GM130, red. Scale bars, 5 μm.
From: Lopes-da-Silva M, McCormack JJ, Burden JJ, Harrison-Lavoie KJ, Ferraro F, Cutler DF.
A GBF1-Dependent Mechanism for Environmentally Responsive Regulation of ER-Golgi Transport.
Dev Cell. 2019 Jun 3;49(5):786-801.e6.
doi: 10.1016/j.devcel.2019.04.006.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to label the trans-golgi network in HeLa cells by immunofluorescence.
Image caption:
OROV recruits Golgi complex and TGN proteins to its assembly site.
HeLa cells were infected with OROV and fixed after 0 h (A-D) or 24 h p.i. (F-I). Golgi complex and TGN proteins (Giantin and TGN46, respectively) were co-immunostained with OROV proteins and analyzed by confocal microscopy. Bars = 10 μm. (E and J) Insets representing the boxed areas of A-D and F-I, respectively. Bars = 2 μm.
From: Barbosa NS, Mendonça LR, Dias MVS, Pontelli MC, da Silva EZM, Criado MF, da Silva-Januário ME, Schindler M, Jamur MC, Oliver C, Arruda E, daSilva LLP.
ESCRT machinery components are required for Orthobunyavirus particle production in Golgi compartments.
PLoS Pathog. 2018 May 3;14(5):e1007047.
doi: 10.1371/journal.ppat.1007047.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to demonstrate the trans golgi network by immunofluorescence.
Image caption:
ηCTF fragment is localized in Golgi and endosomes. a–d Hela cells were transiently transfected with ηCTF and immunostained with ηCTF-Nter (green) antibodies for ηCTF detection and antibodies directed towards TGN-46 (trans-Golgi apparatus, red (a),
From: Afram E, Lauritzen I, Bourgeois A, El Manaa W, Duplan E, Chami M, Valverde A, Charlotte B, Pardossi-Piquard R, Checler F.
The η-secretase-derived APP fragment ηCTF is localized in Golgi, endosomes and extracellular vesicles and contributes to Aβ production.
Cell Mol Life Sci. 2023 Mar 17;80(4):97.
doi: 10.1007/s00018-023-04737-4.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500) used to label the trans golgi network in HeLa cells by immunofluorescence
Image caption:
Subcellular localization of EGFP-SCOC (isoform 3). B. HeLa Flp-In T-Rex cells stably expressing EGFP-SCOC (isoform 3. WT) or EGFP-SCOC (isoform 3, F14A/I17A) starved for 2 h in EBSS, fixed and labelled with anti-TGN46 and Hoechst for confocal microscopy. Expression of EGFP-SCOC constructs was induced with 0.5 μg/ml tetracycline overnight. Scale bars represent 10 μM.
From: Wirth M, Mouilleron S, Zhang W, Sjøttem E, Princely Abudu Y, Jain A, Lauritz Olsvik H, Bruun JA, Razi M, Jefferies HBJ, Lee R, Joshi D, O'Reilly N, Johansen T, Tooze SA.
Phosphorylation of the LIR Domain of SCOC Modulates ATG8 Binding Affinity and Specificity.
J Mol Biol. 2021 Jun 25;433(13):166987.
doi: 10.1016/j.jmb.2021.166987.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to evaluate TGN46 expression in HeLa cell lysates by Western blotting.
Image caption:
TGN46 is required for cargo sorting and loading into CARTS.
(A) Pancreatic adenocarcinoma upregulated factor (PAUF) secretion assay in parental (WT) and CRISPR/Cas9-mediated TGN46 knockout (KO) HeLa cells. Cells expressing PAUF-MycHis were incubated for 4 hr in fresh medium without serum in the absence (Ctrl) or presence of Brefeldin A (BFA), after which the cells were lysed. Equal amounts of cell lysates and their corresponding secreted fraction (medium) were analyzed by Western blotting (left) using an anti-Myc antibody (top blot) and anti-TGN46 antibody (bottom blot). BFA was included as a positive control of secretion inhibition.
From: Lujan P, Garcia-Cabau C, Wakana Y, Vera Lillo J, Rodilla-Ramírez C, Sugiura H, Malhotra V, Salvatella X, Garcia-Parajo MF, Campelo F.
Sorting of secretory proteins at the trans-Golgi network by human TGN46.
Elife. 2024 Mar 11;12:RP91708.
doi: 10.7554/eLife.91708.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to evaluate TGN46 expression in HeLa cell lysates by immunofluorescence..
Image caption:
TGN46 is required for cargo sorting and loading into CARTS.
(E) Left panels are confocal fluorescence microscopy images of HeLa cells transfected with control (Ctrl) or TGN46 siRNA, which were also transfected with PKD2-KD-Flag and PAUF-mRFP. Cells were fixed at steady state and labeled with anti-TGN46 (cyan) and anti-Flag (magenta) antibodies. PAUF-mRFP fluorescence signal is shown in green. Scale bars are 10 μm, magnifications of the boxed regions are shown.
From: Lujan P, Garcia-Cabau C, Wakana Y, Vera Lillo J, Rodilla-Ramírez C, Sugiura H, Malhotra V, Salvatella X, Garcia-Parajo MF, Campelo F.
Sorting of secretory proteins at the trans-Golgi network by human TGN46.
Elife. 2024 Mar 11;12:RP91708.
doi: 10.7554/eLife.91708.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label the trans Golgi network in HeLa cells by immunofluorescence.
Image caption:
The Rab27a downstream effector Myosin Va is important for OROV trafficking and release.
HeLa shControl (upper panels) and shMyoVa (lower panels) cells growing on coverslips were inoculated with OROV (MOI = 4) and fixed at 24 h.p.i. Cells were permeabilized and stained with primary anti-OROV (mouse), anti-MyoVa (Rabbit) and anti-TGN46 (sheep) antibodies, followed by secondary anti-mouse IgG 488 (green), anti-rabbit IgG 594 (red) and anti-sheep IgG 647 (cyan). Nuclei are stained with DAPI (in blue). Scale bar = 10 μm. Insets represent the boxed areas. Scale bar = 2 μm. A maximum intensity projection image across the Z-stack for OROV staining is shown in each case. Scale bar = 10 μm.
From: Concha JO, Gutierrez K, Barbosa N, Rodrigues RL, de Carvalho AN, Tavares LA, et al. (2024)
Rab27a GTPase and its effector Myosin Va are host factors required for efficient Oropouche virus cell egress.
PLoS Pathog 20(8): e1012504.
doi: 10.1371/journal.ppat.1012504.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label the trans Golgi network in fibroblasts by immunofluorescence.
Image caption:
MyoVa KO Fibroblasts show alteration in the viral compartments’ morphology and a reduction in OROV release.
MyoVa WT and MyoVa KO fibroblasts growing on cover slips were infected with OROV (MOI = 4). After 24 h, the cells were fixed, permeabilized and incubated with anti-OROV (mouse) and anti-TGN46 (sheep) antibodies, followed anti-mouse IgG 488 (green) and anti-sheep IgG 594 (red) secondary antibodies. Nuclei are stained with DAPI (in blue). Scale bar = 10 μm. Insets represent the boxed areas. Scale bar = 2 μm.
From: Concha JO, Gutierrez K, Barbosa N, Rodrigues RL, de Carvalho AN, Tavares LA, et al. (2024)
Rab27a GTPase and its effector Myosin Va are host factors required for efficient Oropouche virus cell egress.
PLoS Pathog 20(8): e1012504.
doi: 10.1371/journal.ppat.1012504.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label the trans Golgi network in HeLa cells by immunofluorescence.
Image caption:
Rab27a colocalizes with OROV in late stages of infection.
HeLa shControl (upper panels) and shRab27a (lower panels) cells growing on coverslips were inoculated with OROV (MOI = 4) and fixed at 24 hours post-infection. Cells were permeabilized and stained with primary mouse anti-OROV, rabbit anti-Rab27a and sheep anti-TGN46 antibodies, followed by secondary anti-mouse IgG 488 (green), anti-rabbit IgG 594 (red) and anti-sheep IgG 647 (cyan). Nuclei are stained with DAPI (in blue). Scale bar = 10 μm. Insets represent the boxed areas. Scale bar = 2 μm. A maximum intensity projection image across the Z-stack for OROV staining is shown in each case. Scale bar = 10 μm.
From: Concha JO, Gutierrez K, Barbosa N, Rodrigues RL, de Carvalho AN, Tavares LA, et al. (2024)
Rab27a GTPase and its effector Myosin Va are host factors required for efficient Oropouche virus cell egress.
PLoS Pathog 20(8): e1012504.
doi: 10.1371/journal.ppat.1012504.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46 antibody (AHP500G) used to label the trans Golgi network in HeLa cells by immunofluorescence.
Image caption:
Overexpression of Rab27a mutants affect OROV intracellular distribution and release.
(E) HeLa cell lines with inducible expression of Rab27a WT, Rab27a Q78L or Rab27a T23N mutants, growing on cover slips were infected with OROV (MOI = 4) and after 4 hours the cells were treated with 10ng/ml of Doxycycline to induce the expression. After 24 h.p.i., the cells were fixed, permeabilized and incubated with anti-OROV (mouse), anti-Rab27a (rabbit) and anti-TGN46 (sheep) antibodies, followed anti-mouse IgG 488 (green), anti-rabbit IgG 594 (red) and anti-sheep IgG 647 (cyan) secondary antibodies. Nuclei are stained with DAPI (in blue). Scale bar = 10 μm. (F) Colocalization between TGN46 and OROV from (E), bars represent the mean ± SD of the Manders’ colocalization coefficient (at least 10 cells from three independent experiments). p>0.05 was considered as not significant (one-way ANOVA followed by Tukey´s multiple comparisons test).
From: Concha JO, Gutierrez K, Barbosa N, Rodrigues RL, de Carvalho AN, Tavares LA, et al. (2024)
Rab27a GTPase and its effector Myosin Va are host factors required for efficient Oropouche virus cell egress.
PLoS Pathog 20(8): e1012504.
doi: 10.1371/journal.ppat.1012504.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Human TGN46
- Product Type
- Polyclonal Antibody
- Isotype
- Polyclonal IgG
- Specificity
- TGN46
Antibody Quality and Performance Guarantee
| Sheep anti Human TGN46 antibody recognizes trans-Golgi network integral membrane protein 2 (TGOLN2), also known as TGN38 homolog, TGN46, TGN48 or trans-Golgi network protein TGN51. TGN46 is a 437 amino acid glycoprotein localized to the trans-Golgi network. TGN46 has been reported as being the best available marker for human trans-Golgi network. TGN46 is a heavily glycosylated protein of around 110-120 kDa. Multiple isoforms of TGN46 are generated by alternative splicing differing in sequence at the C-terminal portion. Sheep anti Human TGN46 antibody is expected to recognize all identified isoforms. |
- Target Species
- Human
- Species Cross-Reactivity
-
Target Species Cross Reactivity Primate - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Antiserum Preparation
- Antisera to human TGN46 were raised by repeated immunisation of sheep with highly purified antigen. Purified IgG prepared by affinity chromatography.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
0.5% Bovine Serum Albumin
25% Glycerol - Immunogen
- Recombinant human TGN46.
- Approx. Protein Concentrations
- IgG concentration 0.25 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Immunofluorescence |  |
||
| Immunohistology - Frozen 1 | |
0.1ug/ml | 1ug/ml |
| Western Blotting | |
0.1ug/ml | 1ug/ml |
- 1Fixation with 3% paraformaldehyde or methanol/acetone is recommended.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Rabbit anti Sheep IgG (H/L):Biotin | 5184-2304 | C E F WB | 1.5 ml |
|
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| List Price | Your Price | ||||||
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| Description | Rabbit anti Sheep IgG (H/L):Biotin | ||||||
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Further Reading
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Ponnambalam, S. et al. (1996) Primate homologues of rat TGN38: primary structure, expression and functional implications.
J Cell Sci. 109 ( Pt 3): 675-85.
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