|Immunofluorescence (Frozen)||1:50 - 1:100|
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Cover sections with 4% formaldehyde dilute in 1X PBS.
NOTE: Formaldehyde is toxic, use only in fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 222
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of mouse CD20 protein.
B-lymphocyte antigen CD20 (also known as MS4A1; Membrane-spanning 4-domains subfamily A member 1) is a cell surface phosphoprotein involved in the regulation of B cell activation and proliferation (1,2). It is commonly used as a marker to identify B cells and is expressed throughout B cell development, up until their differentiation into plasma cells. CD20 has no known ligand, and its expression and function are largely conserved between human and mouse (1-3). Evidence suggests that CD20 is necessary for store operated calcium (SOC) entry, which leads to elevated cytoplasmic calcium levels required for B cell activation (4,5). Anti-CD20 antibody immunotherapy depletes B cells by activation of the innate monocytic network and is a common treatment for B cell lymphomas, leukemias, and autoimmune diseases (6).
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