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BUV615 Rat Anti-Mouse CD21/CD35
Product Details
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BD OptiBuild™
CR2/CR1
Mouse (Tested in Development)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Purified Mouse CR1
Flow cytometry (Qualified)
0.2 mg/ml
12902
AB_2875176
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. CF™ is a trademark of Biotium, Inc.
  10. BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
751152 Rev. 2
Antibody Details
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7G6

The 7G6 antibody recognizes an epitope shared by 145-150-kDa and 190-kDa complement receptor proteins, originally designated CR2 (CD21) and CR1 (CD35), respectively.  In the mouse, CD21 and CD35 are expressed on the majority of peripheral B lymphocytes, on the majority of resident peritoneal macrophages and mast cells, 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.  CD21 is a ligand-binding component of the CD19/CD21/CD81 signal-transduction complex associated with the antigen receptor on B lymphocytes.  CD21/CD35 also co-localizes with CD19 on the surface of peritoneal mast cells.   Cr2null mice display impaired inflammatory and humoral immune responses in vivo.  The 7G6 mAb has been reported to inhibit rosette formation by C3d-bearing sheep erythrocytes, to block the complement dependent trapping of immune complexes by follicular dendritic cells, and to down-regulate mouse CD21/CD35 expression upon in vivo application, thus inhibiting primary antibody responses to immunization.  Co-stimulation of B-cell differentiation via Sepharose-coupled 7G6 antibody has also been observed.  The 7G6 mAb recognizes an epitope on CD35 distinct from the epitope recognized by anti-mouse CD35, clone 8C12, and it does not block binding of 8C12 mAb to mouse CD35.

The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP.  Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).

751152 Rev. 2
Format Details
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BUV615
The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV615
Ultraviolet 355 nm
350 nm
615 nm
751152 Rev.2
Citations & References
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Development References (15)

  1. Axcrona K, Gray D, Leanderson T. Regulation of B cell growth and differentiation via CD21 and CD40. Eur J Immunol. 1996; 26(9):2203-2207. (Clone-specific: (Co)-stimulation, Flow cytometry). View Reference
  2. Cariappa A, Tang M, Parng C, et al. The follicular versus marginal zone B lymphocyte cell fate decision is regulated by Aiolos, Btk, and CD21. Immunity. 2001; 14(5):603-615. (Clone-specific: Flow cytometry). View Reference
  3. Fagarasan S, Muramatsu M, Suzuki K, Nagaoka H, Hiai H, Honjo T. Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora. Science. 2002; 298(5597):1424-1427. (Clone-specific: Fluorescence microscopy, Immunofluorescence). View Reference
  4. Fischer MB, Goerg S, Shen L, et al. Dependence of germinal center B cells on expression of CD21/CD35 for survival. Science. 1998; 280(5363):582-585. (Biology). View Reference
  5. Gommerman JL, Oh DY, Zhou X, et al. A role for CD21/CD35 and CD19 in responses to acute septic peritonitis: a potential mechanism for mast cell activation. J Immunol. 2000; 165(12):6915-6921. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
  6. Heyman B, Wiersma EJ, Kinoshita T. In vivo inhibition of the antibody response by a complement receptor-specific monoclonal antibody. J Exp Med. 1990; 172(2):665-668. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
  7. Hu H, Martin BK, Weis JJ, Weis JH. Expression of the murine CD21 gene is regulated by promoter and intronic sequences. J Immunol. 1997; 158(10):4758-4768. (Biology). View Reference
  8. Kinoshita T, Takeda J, Hong K, Kozono H, Sakai H, Inoue K. Monoclonal antibodies to mouse complement receptor type 1 (CR1). Their use in a distribution study showing that mouse erythrocytes and platelets are CR1-negative. J Immunol. 1988; 140(9):3066-3072. (Immunogen: Immunoprecipitation, Radioimmunoassay). View Reference
  9. Kinoshita T, Thyphronitis G, Tsokos GC, et al. Characterization of murine complement receptor type 2 and its immunological cross-reactivity with type 1 receptor. Int Immunol. 1990; 2(7):651-659. (Clone-specific). View Reference
  10. Molina H, Holers VM, Li B, et al. Markedly impaired humoral immune response in mice deficient in complement receptors 1 and 2. Proc Natl Acad Sci U S A. 1996; 93(8):3357-3361. (Clone-specific: Flow cytometry). View Reference
  11. Oliver AM, Martin F, Gartland GL, Carter RH, Kearney JF. Marginal zone B cells exhibit unique activation, proliferative and immunoglobulin secretory responses. Eur J Immunol. 1997; 27(9):2366-2374. (Clone-specific: Flow cytometry). View Reference
  12. Oliver AM, Martin F, Kearney JF. IgMhighCD21high lymphocytes enriched in the splenic marginal zone generate effector cells more rapidly than the bulk of follicular B cells. J Immunol. 1999; 162(12):7198-7207. (Clone-specific: Flow cytometry). View Reference
  13. Thyphronitis G, Kinoshita T, Inoue K, et al. Modulation of mouse complement receptors 1 and 2 suppresses antibody responses in vivo. J Immunol. 1991; 147(1):224-230. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
  14. Wiersma EJ, Kinoshita T, Heyman B. Inhibition of immunological memory and T-independent humoral responses by monoclonal antibodies specific for murine complement receptors. Eur J Immunol. 1991; 21(10):2501-2506. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
  15. Yoshida K, van den Berg TK, Dijkstra CD. Two functionally different follicular dendritic cells in secondary lymphoid follicles of mouse spleen, as revealed by CR1/2 and FcR gamma II-mediated immune-complex trapping. Immunology. 1993; 80(1):34-39. (Clone-specific: Inhibition). View Reference
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751152 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.