CD20 Monoclonal Antibody (2H7), APC-eFluor™ 780, eBioscience™, Invitrogen™

Description

Description: The 2H7 monoclonal antibody reacts with human CD20, a 33-36 kDa transmembrane protein. CD20 is expressed by developing B cells as well as mature B cells but not plasma cells. CD20 has been detected at low levels on a small subset of mature T cells. It is suggested that CD20 plays a role in B-cell activation. Applications Reported: This 2H7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 2H7 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blod cells. This can be used at 5 μL (0.25 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. APC-eFluor 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL cell sample + 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.

CD20 is a non-glycosylated surface phosphoprotein that has a molecular weight range of 33-37 kDa depending on the degree of phosphorylation. CD20 is expressed on mature and most malignant B cells, in a subpopulation of T lymphocytes and follicular dendritic cells. CD20 expression on B cells is synchronous with the expression of surface IgM and it regulates transmembrane calcium conductance, cell cycle progression and B-cell proliferation. CD20 is also associated with lipid rafts, but the intensity of this association depends on extracellular triggering, employing CD20 conformational change, and/or BCR (B cell antigen receptor) aggregation. After the receptor ligation, BCR and CD20 colocalize and then rapidly dissociate before BCR endocytosis, whereas CD20 remains at the cell surface. CD20 serves as a useful target for antibody-mediated therapeutic depletion of B cells, as it is expressed at high levels on most B-cell malignancies, but does not become internalized or shed from the plasma membrane following monoclonal antibody treatment. Diseases associated with CD20 dysfunction include Ms4a1-related common variable immune deficiency.