CD63 Monoclonal Antibody (H5C6), PE-eFluor™ 610, eBioscience™

Product Specific Information

Description: This H5C6 monoclonal antibody reacts with human CD63, a type III member of the tetraspanin family of transmembrane proteins. CD63 is expressed intracellularly on lysosomes, endosomes, and granules of resting platelets and basophils. However, cell surface expression of CD63 can be detected on activated basophils and platelets, monocytes, macrophages, and granulocytes. This receptor is also expressed on endothelial cells, fibroblasts, and smooth muscle cells. Studies have demonstrated that CD63 associates with integrins (VLA-3 and VLA-6) and TIMP-1 to mediate the allergic response.

Applications Reported: This H5C6 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This H5C6 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (1.0 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

PE-eFluor 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red). Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Target Information

CD63, also known as LAMP-3, is a glycoprotein belonging to the tetraspanin family, primarily located in late endosomes, lysosomes, and secretory vesicles across various cell types. It is expressed intracellularly in resting platelets and basophils, but upon activation, CD63 translocates to the plasma membrane, making it a widely used marker for basophil activation. Unlike basophils, mast cells can expose CD63 without activation. CD63 is expressed on activated platelets, monocytes, macrophages, and granulocytes, as well as endothelial cells, fibroblasts, and smooth muscle cells. It interacts with integrins (such as VLA-3 and VLA-6) and TIMP-1, playing roles in mediating allergic responses, phagocytosis, and cell migration. Additionally, CD63 is involved in the regulation of H/K-ATPase trafficking and ROMK1 channels, and serves as a T-cell costimulation molecule. CD63 expression can be used to predict prognosis in early-stage carcinomas and is identical to the melanoma-associated antigen ME491 and the platelet antigen PTLGP40. Diseases associated with CD63 dysfunction include melanoma and Hermansky-Pudlak Syndrome, highlighting its significance in both immune responses and disease processes.