TGN46 Antibody

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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 PKCd 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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TGN46 Antibody gallery image 19

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Sheep anti Human TGN46 antibody 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.

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Published customer image:
Sheep anti Human TGN46 antibody 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.

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TGN46 Antibody gallery image 24

Published customer image:
Sheep anti Human TGN46 antibody 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.

Enlarge
TGN46 Antibody gallery image 25

Published customer image:
Sheep anti Human TGN46 antibody 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.

Enlarge
TGN46 Antibody gallery image 26

Published customer image:
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 5&MUm. (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.

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  • Sheep anti Human TGN46
  • Sheep anti Human TGN46
  • Sheep anti Human TGN46
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Polyclonal Antibody
  • Isotype
    Polyclonal IgG
2 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    AHP500C, IF, WBdatasheet pdfdatasheet pdf0.1 ml
    AHP500
    AHP500GTC *, IF, WBdatasheet pdfdatasheet pdf10 µg
    AHP500GT
    AHP500GC *, IF, WBdatasheet pdfdatasheet pdf25 µg
    AHP500G
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • 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 difffering in sequence at the C-terminal protion. Sheep anti Human TGN46 antibody is expeced to recognize all identified isoforms.
    • Intended Use
    • Target Species
      Human
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Primateyes
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Serum, diluted - liquid
      Purified IgG - liquid
      Purified IgG - liquid
    • Reconstitution
    • Preparation
    • Antiserum Preparation
      Antisera to human TGN46 were raised by repeated immunisation of sheep with highly purified antigen. Purified IgG prepared by affinity chromatography.
    • Preservative Stabilisers
      0.02% Sodium Azide (NaN3)
      1% Bovine Serum Albumin
      25% Glycerol
      Pack Size: 25 µg0.09% Sodium Azide (NaN3)
      0.5% Bovine Serum Albumin
      25% Glycerol
      Pack Size: 10 µg0.09% Sodium Azide (NaN3)
      1% Bovine Serum Albumin
      25% Glycerol
      Pack Size: 25 µg0.09% Sodium Azide (NaN3)
      0.5% Bovine Serum Albumin
      25% Glycerol
      Pack Size: 10 µg0.09% Sodium Azide (NaN3)
      1% Bovine Serum Albumin
      25% Glycerol
    • Immunogen
      Recombinant human TGN46.
    • Purity
    • Approx. Protein Concentrations
      Pack Size: 25 µgIgG concentration 0.25 mg/ml
      Pack Size: 10 µgIgG concentration 0.25mg/ml
      Pack Size: 25 µgIgG concentration 0.25 mg/ml
      Pack Size: 10 µgIgG concentration 0.25mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.

      This product should be stored undiluted.

      Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.

      This product should be stored undiluted.

      Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.

      This product should be stored undiluted.

      Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
      18 months from date of despatch.
      18 months from date of despatch.
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • 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 NameYesNoMin DilutionMax Dilution
      Immunofluorescence
      Immunohistology - Frozen1/501/100
      Western Blotting1/5001/000
    • Application NameYesNoMin DilutionMax Dilution
      Immunofluorescence
      Western Blotting0.1ug/ml1ug/ml
      Immunohistology - Frozen(1)0.1ug/ml1ug/ml
      (1)
      Fixation with 3% paraformaldehyde or methanol/acetone is recommended.
    • Application NameYesNoMin DilutionMax Dilution
      Immunofluorescence
      Western Blotting0.1ug/ml1ug/ml
      Immunohistology - Frozen(1)0.1ug/ml1ug/ml
      (1)
      Fixation 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.
    • Technical Advice
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • Recommended Protocol
    • ELISA
    • ELISA
    • ELISA
    • Immunohistology
      Fixation with 3% paraformaldehyde or methanol/acetone is recommended.
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
    • Western Blotting
    • Instructions For Use
    • Instructions For Use
    • Instructions For Use

    Additional TGN46 Antibody Formats

    Formats Applications Sizes available
    TGN46 Antibody : Serum C, IF, WB 0.1 ml
    TGN46 Antibody : Purified C *, IF, WB 10 µg | 25 µg
    • Copyright © 2016 Bio-Rad

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      Donkey anti Sheep/Goat IgG:DyLight®488STAR88D488GA0.1 mgF, IF
      STAR88D488GA
      Donkey anti Sheep/Goat IgG:DyLight®549STAR88D549GA0.1 mgF, IF
      STAR88D549GA
      Donkey anti Sheep/Goat IgG:DyLight®649STAR88D649GA0.1 mgF, IF
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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              • Application NameReference Images
                Immunofluorescence
                Western Blotting

              References

              Further Reading

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