Description
The 8C12 antibody recognizes an epitope present on the 190-kDa complement receptor protein, originally designated CR1 (CD35), but not the 145-150-kDa CR2 (CD21) molecule. Unlike the human system, in which these proteins are products of independent genes, both of these mouse receptors are membrane proteins resulting from the alternative splicing of mRNA transcribed from the Cr2 gene. Therefore, an alternative nomenclature has been proposed, designating the proteins Cr2-190 (CD21b) and Cr2-145 (CD21a), respectively. The epitope recognized by 8C12 mAb is only present on CD35/CD21b. Moreover, it has also been proposed that Crry is the true mouse genetic homologue of human CR1 (CD35). In the mouse, CD35 is expressed on the majority of peripheral B cells, on the majority of resident peritoneal macrophages, on peripheral blood granulocytes after treatment with N-formyl-Met-Leu-Phe, and on follicular dendritic cells, but not on thymocytes, T cells, erythrocytes, or platelets. In addition, it has not been detected, at the protein or mRNA level, in the macrophage cell line J774, bone marrow-derived macrophages, or thioglycollate-elicited peritoneal macrophages. The 8C12 mAb has been reported to inhibit rosette formation by C3bbearing sheep erythrocytes, to block the complement-dependent trapping of immune complexes by follicular dendritic cells, and to down-regulate mouse CD35 expression upon in vivo application, inhibiting only some primary antibody responses to immunization. B lymphocytes of Cr2[null] mice display impaired humoral immune responses in vivo. The 8C12 mAb recognizes an epitope on mouse CD35 distinct from the epitope recognized by anti-mouse CD21/CD35 mAb 7G6, and it does not block binding by 7G6 mAb to CD35. *Please note that the isotype of 8C12 mAb was originally reported to be Rat IgG2c. Further investigations have demonstrated that the isotype of 8C12 mAb is Rat IgG2a.
Put all BD® AbSeq Reagents to be pooled into a Latch Rack for 500 µL Tubes (Thermo Fisher Scientific Cat. No. 4900). Arrange the tubes so that they can be easily uncapped and re-capped with an 8-Channel Screw Cap Tube Capper (Thermo Fisher Scientific Cat. No. 4105MAT) and the reagents aliquoted with a multi-channel pipette. BD® AbSeq tubes should be centrifuged for ≥ 30 seconds at 400 × g to ensure removal of any content in the cap/tube threads prior to the first opening.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography and conjugated to BD® AbSeq oligonucleotide under optimal conditions.
1. This reagent has been pre-diluted for use at the recommended volume per test. Typical use is 2 µl for 1 × 10^6 cells in a 200-µl staining reaction.
2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
3. Please refer to bd.com/genomics-resources for technical protocols.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
8. For U.S. patents that may apply, see bd.com/patents.
