CD63 antibody | MEM-259
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to evaluate CD63 expression on erythroid cells in vitro by flow cytometry.
Image caption:
Tetraspanin and integrin profile of erythroid cultures. Results are depicted from one culture where directly conjugated antibodies were used (days 3 and 4) and a second culture with indirectly labelled antibodies (day 5 onwards). The y-axis scale is linear to 350 counts; the x-axis is logarithmic to 104. Images were captured on a Leica DM750 microscope, x20 magnification, using Image-Pro Express 6.0 software.
From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, Chasis JA, et al. (2013)
Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1.
PLoS ONE 8(5): e62654.
doi: 10.1371/journal.pone.0062654
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to evaluate CD63 expression on erythroid cells in vitro by immunofluorescence.
Image caption:
Confocal imaging of erythroblasts.
(A) Dual staining of α4 (Red) and β1 (Red) with tetraspanin CD63 (green) on days 5, 8 and 12 of culture. Colocalisation is seen in yellow on the coloured images and the adjacent grayscale images highlight yellow areas of colocalisation only (scale bars = 10 μm).
From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, Chasis JA, et al. (2013)
Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1.
PLoS ONE 8(5): e62654.
doi: 10.1371/journal.pone.0062654
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to evaluate CD63 expression on erythroid cells in vitro by immunofluorescence.
Image caption:
Confocal imaging of reticulocytes.
Dual staining of tetraspanin CD63 (green) with α4 and β1 integrins (both red) on day 12 of culture shown in phase contrast and fluorescence (scale bars = 10 μm). Colocalisation is seen in yellow. (B) Single examples of reticulocytes zoomed in from images in (A) shown in phase contrast, fluorescence and greyscale images. Yellow on phase contrast and fluorescence denotes colocalisation. The righthand grayscale images highlight yellow areas of colocalisation only (scale bars = 5 μm).
From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, Chasis JA, et al. (2013)
Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1.
PLoS ONE 8(5): e62654.
doi: 10.1371/journal.pone.0062654
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to evaluate CD63 expression on erythroid cells in vitro by immunoprecipitation and western blotting.
Image caption:
Tetraspanins CD81, CD82 and CD151 are associated with α4β1 throughout erythroid maturation and with β3 in proerythroblasts and basophilic erythroblasts.
B. Tetraspanin precipitates from day 6 proerythroblasts (ProEB) solubilised in the presence of EDTA or different cations, and from Mn2+-activated basophilic erythroblasts (BasoEB, day 8) were probed with a mix of antibodies to α5, β1, β2 and β3 integrins while the control samples were probed with the relevant tetraspanin antibodies. For clarity, integrin controls are illustrated for the EDTA blot but were present on all blots. β1 and β3 integrins were precipitated well only in the presence of Mn2+.
Each tetraspanin co-precipitates the other most strongly in the presence of Mn2+ from proerythroblasts while any cation permits co-precipitation in HEL cells. Integrins were analysed on 7.5% gels, tetraspanins on 12% gels; non-reducing conditions. Unless stated, the following clones were used: CD53, MEM-53; CD63, MEM-259; CD81, 454720; CD82, TS82b; CD151, IIG5a; α4, HP2/1; α5, IIA1; αL, TS1/22; αIIb, PAB-1. All day 5 and 6 cultures comprised 90–95% proerythroblasts; day 8 culture comprised 5% proerythroblasts, 81% basophilic erythroblasts and 14% polychromatic erythroblasts; day 12 culture comprised 41% polychromatic erythroblasts, 15% orthochromatic erythroblasts and 41% reticulocytes. In the day 5 and 6 cultures 15–34% of cells were GPA+ and 28–35% of cells were αIIb+. Day 8 and day 12 cultures had 77% and 97% GPA+ cells, respectively, and 9% and 0% αIIb+ cells, respectively.
From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, Chasis JA, et al. (2013)
Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1.
PLoS ONE 8(5): e62654.
doi: 10.1371/journal.pone.0062654
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to evaluate CD63 expression on erythroid cells in vitro by immunoprecipitation.
Image caption:
Co-precipitation of β1 and β3 integrins by tetraspanins with different cations from HEL cells. Precipitates were prepared from HEL cells solubilised in Brij-97 in the presence of EDTA or cations. β1 is co-precipitated by CD63, CD81, CD82 and CD151 while β3 is co-precipitated by all tetraspanins in the presence of Mn2+. CD151 co-precipitates β1 under all conditions. Integrins were separated on 7.5% gels, tetraspanin controls on 12% gels, both non-reducing conditions. CD53, MEM-53; CD63, MEM-259; CD81, 454720; CD82, TS82b; CD151, IIG5a; α4, HP2/1; αL, TS1/22; αIIb, PAB-1.
From: Spring FA, Griffiths RE, Mankelow TJ, Agnew C, Parsons SF, Chasis JA, et al. (2013)
Tetraspanins CD81 and CD82 Facilitate α4β1-Mediated Adhesion of Human Erythroblasts to Vascular Cell Adhesion Molecule-1.
PLoS ONE 8(5): e62654.
doi: 10.1371/journal.pone.0062654
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to indicate the distribution of CD63 in relation to LAMP-1 in SH-SY5Y neuroblastoma cells cultured in the presence of β2m fibrils by immunofluorescence.
Image caption:
Fragmented fibrils of β2m increase the cell surface expression of LAMP-1 and CD63. SH-SY5Y cells were incubated with 1.2 μm (monomer-equivalent concentration) fragmented β2m fibrils or β2m monomers for 2 and 24 h. A, cells were fixed, permeabilized, and stained with LAMP1 and CD63 antibodies. Antibody staining was detected with fluorescently labeled secondary antibodies. Cell-associated fluorescence was visualized by confocal microscopy. In the merged images, images of antibody staining were combined with phase-contrast images. Yellow indicates colocalization of CD63 (green) and LAMP-1 (red). Scale bar = 10 μm.
From: Jakhria T, Hellewell AL, Porter MY, Jackson MP, Tipping KW, Xue WF, Radford SE, Hewitt EW. β2-microglobulin amyloid fibrils are nanoparticles that disrupt lysosomal membrane protein trafficking and inhibit protein degradation by lysosomes.
J Biol Chem. 2014 Dec 26;289(52):35781-94.
doi: 10.1074/jbc.M114.586222.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Mouse anti Human CD63 antibody, clone MEM-259 (MCA2142) used to localize CD63 expression in HeLa cells by immunofluorescence.
Image caption:
Apilimod induces TGF-βR2 mislocalization to late-endosomes. (A) Serum-starved HeLa cells expressing flag-TGFβ-R2 were treated as indicated with Apilimod or DMSO, fixed and stained with an anti-flag antibody (green) and an anti-CD63 antibody (red). Bar is 10μm.
From: Cinato M, Guitou L, Saidi A, Timotin A, Sperazza E, Duparc T, Zolov SN, Giridharan SSP, Weisman LS, Martinez LO, Roncalli J, Kunduzova O, Tronchere H, Boal F.
Apilimod alters TGFβ signaling pathway and prevents cardiac fibrotic remodeling.
Theranostics. 2021 Apr 19;11(13):6491-506.
doi: 10.7150/thno.55821.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Human CD63
- Product Type
- Monoclonal Antibody
- Clone
- MEM-259
- Isotype
- IgG1
- Specificity
- CD63
Antibody Quality and Performance Guarantee
| Mouse anti Human CD63 antibody, clone MEM-259 recognizes the CD63 cell surface antigen, also known as granulophysin, lysozomal associated membrane protein 3 (LAMP-3), Melanoma-associated antigen ME491, Ocular melanoma-associated antigen or tetraspanin-30. CD63 is a 238 amino acid, tetraspanin glycoprotein of ~40-60 kDa. CD63 is expressed by monocytes, macrophages, endothelial cells where it is a component of Weibel-Palade bodies (Vischer et al. 1993) and by a variety of cell lines. It is located intracellularly in lysosomal granules of platelets, being translocated to the surface upon activation. It may therefore by useful as a marker of platelet activation (Hamamoto et al. 1994). CD63 expression is also associated with melanoma development (Radford et al. 1997) and expression is rapidly down-regulated by progesterone in endometrial tissues (Okada et al. 1999). |
- Target Species
- Human
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% sodium azide (NaN3)
- Immunogen
- HPB-ALL cell line.
- Approx. Protein Concentrations
- IgG concentration 1.0 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 |
|---|---|---|---|
| Flow Cytometry |  |
1/25 | 1/100 |
| Immunofluorescence | |
||
| Immunoprecipitation | |
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.
- Flow Cytometry
- Use 10μl of the suggested working dilution to label 106 cells in 100μl or 100μl whole blood
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Mouse IgG1 Negative Control | MCA928 | F | 100 Tests |
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| List Price | Your Price | ||||||
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| Description | Mouse IgG1 Negative Control | ||||||
References for CD63 antibody
-
Casey, T.M. et al. (2007) Organelle proteomics: identification of the exocytic machinery associated with the natural killer cell secretory lysosome.
Mol Cell Proteomics. 6 (5): 767-80. -
Durand-Panteix, S. et al. (2012) B7-H1, which represses EBV-immortalized B cell killing by autologous T and NK cells, is oppositely regulated by c-Myc and EBV latency III program at both mRNA and secretory lysosome levels.
J Immunol. 189 (1): 181-90. -
Abdel-Latif, D. et al. (2004) Rac2 is critical for neutrophil primary granule exocytosis.
Blood. 104: 832-9. -
Norling, L.V. et al. (2012) Resolvin D1 limits polymorphonuclear leukocyte recruitment to inflammatory loci: receptor-dependent actions.
Arterioscler Thromb Vasc Biol. 32 (8): 1970-8. -
McKechnie, N.M. et al. (2006) Fas-ligand is stored in secretory lysosomes of ocular barrier epithelia and released with microvesicles.
Exp Eye Res. 83: 304-14. -
Spring, F.A. et al. (2013) Tetraspanins CD81 and CD82 facilitate α4β1-mediated adhesion of human erythroblasts to vascular cell adhesion molecule-1.
PLoS One. 8(5):e62654. -
Pliyev, B.K. (2008) Activated human neutrophils rapidly release the chemotactically active D2D3 form of the urokinase-type plasminogen activator receptor (uPAR/CD87).
Mol Cell Biochem. 321: 111-22. -
Suraeva, N.M. et al. (2015) Changes in the Morphological and Immunological Characteristics of Mel Ibr Melanoma Cells in Response to Chicken Embryo Extract.
Bull Exp Biol Med. 159 (4): 520-3.
View The Latest Product References
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Schäfer, T. et al. (2010) A granular variant of CD63 is a regulator of repeated human mast cell degranulation.
Allergy. 65 (10): 1242-55. -
Jakhria, T. et al. (2014) β2-microglobulin amyloid fibrils are nanoparticles that disrupt lysosomal membrane protein trafficking and inhibit protein degradation by lysosomes.
J Biol Chem. 289 (52): 35781-94. -
Ethier, C. et al. (2016) Calcitriol Reduces Eosinophil Necrosis Which Leads to the Diminished Release of Cytotoxic Granules.
Int Arch Allergy Immunol. 171 (2): 119-29. -
Nielsen, M.H. et al. (2023) Increased extracellular vesicles (EVs) related to T cell-mediated inflammation and vascular function in familial hypercholesterolemia.
Atheroscler Plus. 53: 16-25. -
Ilarraza, R. et al. (2023) Rac2 regulates immune complex-mediated granule polarization and exocytosis in neutrophils.
J Leukoc Biol. 114 (2): 116-125. -
Vrancx, C. et al. (2021) Mechanism of Cellular Formation and In Vivo Seeding Effects of Hexameric β-Amyloid Assemblies.
Mol Neurobiol. 58 (12): 6647-69. -
Gratpain, V. et al. (2024) Influence of a pro-inflammatory stimulus on the miRNA and lipid content of human dental stem cell-derived extracellular vesicles and their impact on microglial activation
Heliyon. : e27025. -
Cinato, M. et al. (2021) Apilimod alters TGFβ signaling pathway and prevents cardiac fibrotic remodeling.
Theranostics. 11 (13): 6491-506.
- RRID
- AB_324562
- UniProt
- P08962
- Entrez Gene
- CD63
- GO Terms
- GO:0002576 platelet degranulation
- GO:0005887 integral to plasma membrane
- GO:0005765 lysosomal membrane
- GO:0030168 platelet activation
- GO:0031088 platelet dense granule membrane
- GO:0031902 late endosome membrane
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