Desmoglein 3 antibody | 5G11
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on human cells by immunofluorescence.
Image caption:
Desmoglein-3 and flotillin-2 colocalise in human epidermis and human epithelial cell lines.
Human skin cryosections, HaCaT and MCF10A cells were immunostained for Dsg3 (red) and flotillin-2 (green). In human epidermis, flotillin-2 is expressed throughout the epidermal layer and colocalises with Dsg3 in keratinocytes of the stratum basale and suprabasale. In HaCaT keratinocytes, flotillin-2 is localised at the cell membrane and in intracellular vesicular structures. Dsg3 is localised at the plasma membrane where it colocalises with flotillin-2. In MCF10A mammary epithelial cells, flotillin-2 and Dsg3 show a more profound colocalisation at the sites of cell-cell contact. The cell lines were grown on glass coverslips for at least 3 days. Scale bar: 20μm.
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on HaCaT cells by immunofluorescence.
Image caption:
Loss of flotillins results in pemphigus vulgaris like localisation of Dsg3 in HaCaT keratinocytes.
(a) HaCaT cells depleted of flotillin-1 (F1-shRNA), flotillin-2 (F2-shRNA) or control shRNA cells (sh-control) were grown for three days on glass coverslips, fixed and stained with Dsg3 (red) and flotillin-2 (green) antibodies. Note that the signals for Dsg3 in flotillin knockdown cells in (a) have been adjusted to make the morphological deficiencies more clear. (b) HaCaT keratinocytes were grown for two days on glass coverslips and treated with 150 μg/ml of either control-IgG (healthy person) or IgG from PV patients for 20 h. Cells were fixed and immunostained for Dsg3 (red) and flotillin-2 (green). Scale bar: 20 μm. (c) Quantification of flotillin-2 localisation in control IgG vs. PV IgG treated cells. At least 8 images/condition (total of >200 cells/condition) originating from three independent experiments were quantified for plasma membrane associated vs. intracellular flotillin-2 staining. Statistical analysis was done by Student's t test (***p <0.001).
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on HaCaT cells by western blotting.
Image caption:
Flotillin depletion results in diminished Dsg3 and E-cadherin protein levels in HaCaT cells.
(a) HaCaT cells with a stable flotillin knockdown or control cells were grown until confluent, and equal protein amounts were analysed by Western blot. (b–f) Western blot signals were quantified by scanning densitometry and normalised against GAPDH. Bars represent the mean ± SD of five (b–e) or four (f) independent experiments. One-way ANOVA with Bonferroni's multiple comparison test. ***p <0.001.
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on HaCaT cells by western blotting.
Image caption:
Flotillin knockdown weakens the adhesion strength in HaCaT keratinocytes but does not affect the raft association or detergent solubility of Dsg1 and Dsg3.
(a) HaCaT cells depleted of flotillin-1 (F1-shRNA) and flotillin-2 (F2-shRNA) and control shRNA cells (sh-control) were dissociated from the plate with dispase and the monolayers were mechanically disrupted. Fragments were fixed, stained and imaged. (b) Relative fragment number from six independent experiments. Bars represent the mean ± SD. One-way ANOVA with Bonferroni's multiple comparison test. *p <0,05, **p <0,01, ***p <0.001. (c–e) Rafts were isolated from control shRNA (c) and flotillin-1 (d) or flotillin-2 (e) depleted HaCaT cells using detergent extraction and density gradient centrifugation. Nine fractions were collected from the top of the gradient (fraction 1 = the lightest fraction) to the bottom and analysed by Western blot. (f) Sequential detergent extraction of HaCaT control and flotillin knockdown cells. Cells were initially extracted with a Triton X-100 buffer followed by solubilisation of the Triton X-100 insoluble proteins with an SDS/urea buffer. Equal volumes of extracted proteins were analysed by Western blot. Please note that the total volume of the insoluble pool is twice that of the soluble one. The lysates shown in the last three rows were extracted with a NP-40 containing lysis buffer to facilitate solubilisation.
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on HaCaT cells by western blotting and immunofluorescence.
Image caption:
Inhibition of lysosomal degradation or endocytosis restores Dsg3 level and improves desmosomal morphology in flotillin knockdown cells.
(a) Flotillin depleted or control HaCaT cells were treated for 24 h with BafA (50 nM) to block lysosomal degradation. Cell lysates were analysed by Western blot. (b) Western blot signals for Dsg3 (upper) and Dsg1 (lower) were quantified by scanning densitometry and normalised against GAPDH. Bars represent the mean ± SD of 6 independent experiments. Two-way ANOVA. *p <0,05, **p <0,01, ***p <0.001. (c) Flotillin knockdown or control cells were serum-starved overnight and treated for two hours with 80 μM Dynasore to inhibit dynamin-dependent endocytic uptake of Dsg3 from the plasma membrane. DMSO served as a control. Cells were fixed and immunostained for Dsg3. Scale bar: 20 μm.
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on human skin cryosections by immunofluorescence.
Image caption:
Desmoglein-3 and flotillin-1 colocalise in human epidermis.
Human skin cryosections were immunostained for Dsg3 (red) and flotillin-1 (green). In human epidermis, flotillin-1 is expressed throughout the epidermal layer and colocalises with Dsg3 in keratinocytes of the stratum basale and suprabasale. In HaCaT keratinocytes, flotillin-1 and flotillin-2 extensively colocalise at the cell membrane and in intracellular vesicular structures.
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human desmoglein-3 antibody, clone 5G11 (MCA2273) used for the evaluation of desmoglein-3 expression on human skin cryosections by immunofluorescence.
Image caption:
Inhibition of lysosomal degradation or endocytosis doesnot change desmoplakin and γ-catenin levels but results in lysosomal accumulation of Dsg3 in flotillin knockdown cells.
Flotillin depleted or control HaCaT cells were treated for24 h with BafA (50 nM) to block lysosomal degradation. After BafA treatment, Dsg3 (red) accumulates in lysosomes and colocalises with the lysosomal marker LAMP1 (green). Scale bar: 20 μm
From: Völlner F, Ali J, Kurrle N, Exner Y, Eming R, Hertl M, Banning A, Tikkanen R.
Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 2016 Jun 27;6:28820.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human Desmoglein 3
- Product Type
- Monoclonal Antibody
- Clone
- 5G11
- Isotype
- IgG1
- Specificity
- Desmoglein 3
Antibody Quality and Performance Guarantee
| Mouse anti Human Desmoglein 3 antibody, clone 5G11 recognizes human desmoglein-3, also known as 130 kDa pemphigus vulgaris antigen or Cadherin family member 6. Desmoglein-3 is a 999 amino acid ~125 kDa single pass type I transmembrane glycoprotein, a core component of the desmosome cell-cell junction between epithelial cells. Desmoglein-3 is synthesized in the basal and lower suprabasal layers of the skin. Mouse anti Human Desmoglein 3 antibody, clone 5G11 does not recognise desmoglein-1 or desmoglein-2. |
- Target Species
- Human
- Product Form
- Purified IgG - liquid
- Preparation
- Antibody purified from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- MCA2273: <0.1%% sodium azide (NaN3)
- MCA2273T: <0.1% Sodium Azide (NaN3)
- Immunogen
- Recombinant human desmoglein 3.
- Approx. Protein Concentrations
- IgG concentration 0.5 mg/ml
- Fusion Partners
- Spleen cells from immunised Balb/c mice were fused with cells of the mouse NS-1 myeloma cell line.
- 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 |  |
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| Immunohistology - Frozen | |
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| Immunohistology - Paraffin 1 | |
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| Immunoprecipitation | |
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| Western Blotting | |
1/100 | 1/1000 |
- 1This product requires protein digestion pre-treatment of paraffin sections e.g. pepsin (Wu et al. 2000). Alternatively it is reported that staining of FFPE material may be enhanced using protein digestion with e.g. pronase, followed by HIER in citrate buffer.
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.
- Histology Positive Control Tissue
- Human skin
- Western Blotting
- Mouse anti human desmoglein 3 antibody, clone 5G11 detects a band of approximately 125 kDa in epithelial cell lysates.
References for Desmoglein 3 antibody
-
Wu, H/ et al. (2000) Protection against pemphigus foliaceus by desmoglein 3 in neonates.
N Engl J Med. 343 (1): 31-5. -
Wahl, J.K. 3rd (2002) Generation of monoclonal antibodies specific for desmoglein family members.
Hybrid Hybridomics. 21 (1): 37-44. -
Wan, H. et al. (2007) Increased keratinocyte proliferation initiated through downregulation of desmoplakin by RNA interference.
Exp Cell Res. 313: 2336-44. -
Szegedi, A. et al. (2008) Protein kinase C isoenzymes differentially regulate the differentiation-dependent expression of adhesion molecules in human epidermal keratinocytes.
Exp Dermatol. 18: 122-9. -
O'Shea, C. et al. (2014) Desmosomal defects in acantholytic squamous cell carcinomas.
J Cutan Pathol. 41 (11): 873-9. -
Völlner, F. et al. (2016) Loss of flotillin expression results in weakened desmosomal adhesion and Pemphigus vulgaris-like localisation of desmoglein-3 in human keratinocytes.
Sci Rep. 6: 28820. -
Kurrle, N. et al. (2013) Flotillins directly interact with γ-catenin and regulate epithelial cell-cell adhesion.
PLoS One. 8 (12): e84393.
Further Reading
-
Koch, P.J. et al. (1998) Desmoglein 3 anchors telogen hair in the follicle.
J Cell Sci. 111 ( Pt 17): 2529-37. -
Jolly, P.S. et al. (2010) p38MAPK signaling and desmoglein-3 internalization are linked events in pemphigus acantholysis.
J Biol Chem. 285: 8936-41.
- RRID
- AB_2246054
- UniProt
- P32926
- Entrez Gene
- DSG3
- GO Terms
- GO:0005509 calcium ion binding
- GO:0016021 integral to membrane
- GO:0005829 cytosol
- GO:0006921 cellular component disassembly involved in apoptosis
- GO:0007156 homophilic cell adhesion
- GO:0030057 desmosome
MCA2273
030615
161161
MCA2273T
160519
180702
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