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Technical Data Sheet

CM0401

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CD63 (Extracellular region)
Mouse Monoclonal IgG2a
Price
Size
Species Reactivity
MW

$295
100 μl
Hu
30-60 kDa

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Western blot of native human cell lyastes: MeWo (lane 1), and MeWo treated with PNGase F (lane 2), and A549 (lane 3), and A549 treated with PNGase F (lane 4). The blot was probed with mouse monoclonal CD63 (CM0401) at 1:100.

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Immunocytochemical labeling of CD63 in paraformaldehyde fixed human MeWo melanoma cells. The cells were labeled with mouse monoclonal CD63 (CM0401). The antibody was detected using goat anti-mouse Ig DyLight® 594.

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Application
Dilution




ICC
1:500


WB
1:100



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Tetraspanins comprise a large superfamily of cell surface-associated membrane proteins with four transmembrane domains. On cell membranes, tetraspanins form networks of various proteins called tetraspanin-enriched microdomains (TEMs). CD63 was the first characterized tetraspanin and it is found in TEMs, as well as late endosomes and lysosomes. In late endosomes, CD63 is enriched on the intraluminal vesicles, and can be secreted as exosomes through fusion of endosomes with the plasma membrane. The complex localization pattern of CD63 suggests that its intracellular trafficking and distribution must be tightly regulated. CD63 contains N-link glycosylation sites that produce diverse CD63 molecules that range from 30 to 60 kDa depending on cell type. CD63 is an important exosomal marker in cancer cells, and may be involved in cancer progression and metastasis.


Background References
Azorsa, D.O. et al. (1991) Blood. 78(2):280.
Metzelaar, M.J. et al. (1991) J Biol Chem. 266(5):3239.
Pols, MS & Klumperman, J (2009) Exp Cell Res. 315(9):1584.
Immunogen
Clone (M040) was generated from a proprietary antigen related to the native human CD63 expressed in MeWo melanoma cell line.
Buffer and Storage
Mouse monoclonal antibody purified with protein G chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
Clone M040 mouse monoclonal antibody has high reactivity to CD63 at the plasma membrane in immunocytochemical assays, and weakly detects a 30-60 kDa* protein on SDS-PAGE "Native" immunoblots of human A431, MeWo, and A549 cells. The antibody does not detect denatured CD63 in western blot assays.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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