Description: The monoclonal antibody QCH6A7 reacts with mouse CD20. CD20 is a B cell specific, 32-kDa membrane protein that is expressed on late pro-B cells through memory B cells in the B cell lineage. CD20 is not found on hematopoietic stem cells, early pro-B cells or later mature plasma cells. CD20 is involved in intracellular calcium regulation and plays a role in B cell proliferation. CD20 is intially expressed after the induction of CD19 during development and is coexpressed with IgM during the pre-B to immature B cell transition in bone marrow. Expression of CD20 increases during maturation with almost all mature B cells expressing some level of CD20. Unlike other B cell antigens, CD20 is not shed or internalized upon antibody binding. Monoclonal antibodies against the CD20 antigen may be used to deplete B cells and CD20 has proven to be an effective immunotheraputic target in B cell lymphomas and several autoimmune diseases including rheumatoid arthritis and type I diabetes. CD20 consists of 4 transmembrane domains and a 44 amino acid extracellular domain which contains the QCH6A7 epitope. Staining with clone QCH6A7 provides significantly improved staining intensity compared to clone AISB12 in flow cytometric assays. Applications Reported: This QCH6A7 antibody has been reported for use in flow cytometric analysis. Applications Tested: This QCH6A7 antibody has been tested by flow cytometric analysis of mouse bone marrow cells and mouse splenocytes.This can be used at less than or equal to 1 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor™ 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. 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 No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 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. Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 μm post-manufacturing filtered. 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 maligt 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 maligcies, 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.
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