Description
The RP/14 monoclonal antibody specifically binds to CD180, the "radioprotective" 105 kDa (RP105) antigen expressed on mature B lymphocytes. Like other members of the Toll-like Receptor (TLR) family, the extracellular region of CD180 contains tandem repeats of a leucine-rich motif flanked by regions containing conserved cysteines. These characteristic structural components of TLR may mediate protein-protein interactions and regulation of signal transduction involved in innate immune responses. Furthermore, CD180 associates with the secretory protein MD-1 on the cell surface. Cross-linking of cell-surface CD180 with the RP/14 antibody induces activation signals in B lymphocytes. Despite the expression of CD180 on lymphocytes of all strains tested (BALB/c, BALB/c.xid, C3H, C57BL/6, CBA/N, and 129/Sv), some strains (BALB/c.xid, C57BL/6.xid, CBA/N, 129/Sv, and 129/Ola) display deficient responses to that stimulatory effects of mAb RP/14.
Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142) may help to reduce non-specific binding to cells bearing Fcγ receptors. If Mouse Fc Block™ is used, then it is important that the second-step anti-rat IgG antibody does not cross-react with the 2.4G2 mAb (rat IgG2b, κ); we recommend the use of biotinylated or FITC-conjugated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894 or 553896, respectively).
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. 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.
4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
