CD20 Monoclonal Antibody (2H7), PE-Cyanine7, 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 blood cells
• This can be used at 5 μL (0.125 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
• Light sensitivity: This tandem dye is sensitive 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
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically
• 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.