Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
Companion Products
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.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.