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Biotin Rat Anti-Mouse CD180
Biotin Rat Anti-Mouse CD180

Two-color analysis of CD180 expression on splenic B lymphocytes. BALB/c splenocytes were stained with either biotinylated rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553928, left panel) or biotinylated mAb RP/14 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by Streptavidin-PE (Cat. No. 554061) and FITC-conjugated anti-mouse CD45R/B220 mAb RA3-6B2 (Cat. No. 553087/553088). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).

Two-color analysis of CD180 expression on splenic B lymphocytes. BALB/c splenocytes were stained with either biotinylated rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553928, left panel) or biotinylated mAb RP/14 (right panel) in the presence of Mouse Fc Block™ (purified anti-mouse CD16/CD32 mAb 2.4G2, Cat. No. 553141/553142), followed by Streptavidin-PE (Cat. No. 554061) and FITC-conjugated anti-mouse CD45R/B220 mAb RA3-6B2 (Cat. No. 553087/553088). Flow cytometry was performed on a FACSCalibur™ (BD Biosciences, San Jose, CA).

Product Details
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BD Pharmingen™
Ly78; Ly-78; Lymphocyte antigen 78; RP105; Radioprotective 105 kDa protein
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
BALB/c mouse splenocytes
Flow cytometry (Routinely Tested)
0.5 mg/ml
17079
AB_394344
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  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.
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Antibody Details
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RP/14

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.

552129 Rev. 1
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
552129 Rev.1
Citations & References
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Development References (7)

  1. 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. (Biology). View Reference
  2. 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. (Biology). View Reference
  3. 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. (Biology). View Reference
  4. 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). View Reference
  5. 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. (Biology). View Reference
  6. 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
  7. 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. (Biology). 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.