Preparation And Storage
Recommended Assay Procedures
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).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
Companion Products
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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.
Development References (7)
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Chan VW, Mecklenbrauker I, Su I, et al. The molecular mechanism of B cell activation by toll-like receptor protein RP-105. J Exp Med. 1998; 188(1):93-101. (Clone-specific: Activation). View Reference
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Corcoran LM, Metcalf D. IL-5 and Rp105 signaling defects in B cells from commonly used 129 mouse substrains. J Immunol. 1999; 163(11):5836-5842. (Clone-specific: Activation). View Reference
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Miyake K, Shimazu R, Kondo J, et al. Mouse MD-1, a molecule that is physically associated with RP105 and positively regulates its expression.. J Immunol. 1998; 161(3):1348-1353. (Clone-specific: Immunoprecipitation). View Reference
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Miyake K, Yamashita Y, Hitoshi Y, Takatsu K, Kimoto M. Murine B cell proliferation and protection from apoptosis with an antibody against a 105-kD molecule: unresponsiveness of X-linked immunodeficient B cells. J Exp Med. 1994; 180(4):1217-1224. (Immunogen: Activation, Apoptosis, Immunoprecipitation). View Reference
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Miyake K, Yamashita Y, Ogata M, Sudo T, Kimoto M. RP105, a novel B cell surface molecule implicated in B cell activation, is a member of the leucine-rich repeat protein family. J Immunol. 1995; 154(7):3333-3340. (Clone-specific: Immunoprecipitation). View Reference
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Ogata H, Su I, Miyake K, et al. The toll-like receptor protein RP105 regulates lipopolysaccharide signaling in B cells. J Exp Med. 2000; 192(1):23-29. (Biology). View Reference
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Yamashita Y, Miyake K, Miura Y, et al. Activation mediated by RP105 but not CD40 makes normal B cells susceptible to anti-IgM-induced apoptosis: a role for Fc receptor coligation. J Exp Med. 1996; 184(1):113-120. (Clone-specific: Activation). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.