Mouse Monoclonal to CD22
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, also known as Siglec-2 (sialic acid-binding immunoglobulin-like lectin-2) is a transmembrane glycoprotein binding alpha2,6-linked sialic acid-bearing ligands. Intracellular domain of CD22 recruits protein tyrosine phosphatase SHP-1 through the immunoreceptor tyrosine-based inhibitory motifs (ITIMs), thus setting a treshold for B cell receptor-mediated activation. CD22 also regulates B-cell response by involvement in controlling the CD19/CD21-Src-family protein tyrosine kinase amplification pathway and CD40 signaling. CD22 exhibits hallmarks of clathrin-mediated endocytic pathway.
The antibody MEM-01 reacts with CD22 (BL-CAM), a 130 kDa type I transmembrane glycoprotein (immunoglobulin superfamily) expressed in the cytoplasm of pro-B and pre-B lymphocytes, and on the surface of mature and activated B lymphocytes; it is lost on plasma cells, peripheral blood T lymphocytes, granulocytes and monocytes.
The antibody MEM-01 cross-blocks the antibody OTH228 that recognizes uniquely epitope "E"; it does not cross-block antibodies RFB-4, CLB22/1 and CLB-BLy1.
Raji Burkitt's lymphoma cell line
- Flow Cytometry
Recommended dilution: 2-4 μg/ml
- Western Blotting
Recommended dilution: 2-4 μg/ml, 120 min on vertical incubator
Positive control: RAJI human lymphoma cell line
Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer.
The antibody MEM-01 stains only the zone corresponding to 140 kDa, but not the weaker 130 kDa zone (stained by several CD22 antibodies recognizing both isoforms).
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.
*Tedder TF, Poe JC, Haas KM: CD22: A Multifunctional Receptor That Regulates B Lymphocyte Survival and Signal Transduction. Adv Immunol. 2005;88:1-50.
*Tateno H, Li H, Schur MJ, Bovin N, Crocker PR, Wakarchuk WW, Paulson JC: Distinct endocytic mechanisms of CD22 (Siglec-2) and Siglec-F reflect roles in cell signaling and innate immunity. Mol Cell Biol. 2007 Aug;27(16):5699-710. [Abstract] [Full Text]
*Walker JA, Smith KG: CD22: an inhibitory enigma. Immunology. 2007 Dec 7 [Abstract]
For research use only. Not for drug, diagnostic or other use.
Mouse IgG1 Isotype Control
Fig. 1. Flow Cytometry analysis
Surface staining of human peripheral blood cells with anti-CD22 (MEM-01
) PE. Cells in the lymphocyte gate were used for analysis.