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BV605 Rat Anti-Mouse CD21/CD35
BV605 Rat Anti-Mouse CD21/CD35
Two-color flow cytometric analysis of CD21/CD35 expression on BALB/c mouse splenocytes. BALB/c splenic leukocytes were simultaneously stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either BD Horizon™ BV605 Rat IgG2b, κ Isotype Control (Cat. No. 563145, Left Panel) or BD Horizon™ BV605 Rat Anti-Mouse CD21/CD35 antibody (Cat. No. 563176, Right Panel) in the presence of Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). Two-color flow cytometric dot plots showing the correlated expression of CD21/CD35 (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD21/CD35 expression on BALB/c mouse splenocytes. BALB/c splenic leukocytes were simultaneously stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880) and either BD Horizon™ BV605 Rat IgG2b, κ Isotype Control (Cat. No. 563145, Left Panel) or BD Horizon™ BV605 Rat Anti-Mouse CD21/CD35 antibody (Cat. No. 563176, Right Panel) in the presence of Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). Two-color flow cytometric dot plots showing the correlated expression of CD21/CD35 (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
CR2/CR1
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Purified Mouse CR1
Flow cytometry (Routinely Tested)
0.2 mg/ml
12902
AB_2738048
Aqueous buffered solution containing BSA and ≤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 with BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  9. CF™ is a trademark of Biotium, Inc.
563176 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.

This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

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).

563176 Rev. 2
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-nm) laser 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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BV605
Violet 405 nm
407 nm
605 nm
563176 Rev.2
Citations & References
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Development References (13)

  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. 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
  12. 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
  13. 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
View All (13) View Less
563176 Rev. 2

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