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      RB670 Mouse Anti-Human CD32
      Product Details
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      BD OptiBuild™
      CD32a/FcγRIIa/FcgRIIA/FCGR2A; CD32b/FcγRIIb/FcgRIIB/FCGR2B
      Human (Tested in Development)
      Mouse BALB/c x A/J, also known as CAF1 IgG2b, κ
      Human K562 leukemia cell line
      Flow cytometry (Qualified)
      0.2 mg/ml
      IV NO89 N504; V MR7
      2212
      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.
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      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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      3. For U.S. patents that may apply, see bd.com/patents.
      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      6. 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.
      7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. An isotype control should be used at the same concentration as the antibody of interest.
      10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      11. Tandem fluorochromes contain both an energy donor and an energy acceptor. 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 the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      12. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      13. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
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      771047 Rev. 1
      Antibody Details
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      IV.3

      Fcγ receptor type II (FcγRII) molecules, also known as CD32 antigens, serve as low affinity receptors for monomeric IgG but bind immune complexes (aggregated IgG) efficiently. Several forms of these receptors, including transmembrane or soluble glycoproteins, are encoded by separate genes and by alternative mRNA splicing: CD32a/FcγRIIa (FCGR2A), CD32b/FcγRIIb (FCGR2B), and CD32c/FcγRIIc (FCGR2C). CD32 molecules are comprised of two IgC-like domains that may be followed by a transmembrane region and a cytoplasmic domain with either ITAM (CD32a and CD32c) or ITIM (CD32b) immunoreceptor signaling motifs. These polymorphic receptors are differentially expressed by leucocyte subsets and are involved in the process of phagocytosis, clearing of immune complexes, platelet activation and degranulation, and regulation of immune responses. The IV.3 monoclonal antibody strongly recognizes FcγRIIA expressed on platelets, monocytes, macrophages, neutrophils, eosinophils, basophils, and B cells. It reportedly binds to an epitope mapped to amino acids 132-137 (FSHLDP) located in the second IgC-like domain within the ligand-binding site. The IV.3 antibody has been used to crosslink or block FcγRII in functional studies as well as a blocking antibody to reduce non-specific binding by antibodies used for staining or cell separation applications. This antibody may weakly crossreact with FcγRIIb/CD32b.

      771047 Rev. 1
      Format Details
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      RB670
      The BD Horizon RealBlue™ 670 (RB670) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492 nm and an emission maximum (Em Max) at 670 nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB670 can be used on both spectral and conventional cytometers and is designed to be primarily excited by the Blue laser (488-nm). For conventional instruments equipped with only a Blue laser (488-nm), RB670 can be used as an alternative to PE-Cy5 and we recommend using an optical filter centered near 670-nm (eg, a 670/30-nm bandpass filter). For conventional and spectral instruments equipped with both a Blue (488-nm) and Yellow-Green (561-nm) laser and appropriate detectors, it can be used in conjunction with PE-Cy5.
      altImg
      RB670
      Blue 488 nm
      492 nm
      670 nm
      771047 Rev.1
      Citations & References
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      View product citations for antibody "771047" on CiteAb

      Development References (11)

      1. Anania JC, Chenoweth AM, Wines BD, Hogarth PM. The Human FcγRII (CD32) Family of Leukocyte FcR in Health and Disease.. Front Immunol. 2019; 10:464. (Biology). View Reference
      2. Boruchov AM, Heller G, Veri MC, Bonvini E, Ravetch JV, Young JW. Activating and inhibitory IgG Fc receptors on human DCs mediate opposing functions.. J Clin Invest. 2005; 115(10):2914-23. (Clone-specific: Flow cytometry). View Reference
      3. Budde P, Weinrich V, Sondermann P, et al. Specificity of CD32 mAb for FcγRIIa, FcγRIIb1, and FcγRIIb2 expressed in transfected mouse B cells and BHK-21 cells. In: Schlossman SF. 1995, ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:828-832.
      4. Fleit HB, Ghazizadeh S. Cross-linking of mAb to FcγRII results in tyrosine phosphorylation of multiple polypeptides including FcγRII itself. In: Schlossman SF. 1995, ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:826-828.
      5. Looney RJ, Abraham GN, Anderson CL. Human monocytes and U937 cells bear two distinct Fc receptors for IgG.. J Immunol. 1986; 136(5):1641-7. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
      6. Looney RJ, Ryan DH, Takahashi K, et al. Identification of a second class of IgG Fc receptors on human neutrophils. A 40 kilodalton molecule also found on eosinophils.. J Exp Med. 1986; 163(4):826-36. (Clone-specific: Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
      7. Rosenfeld SI, Ryan DH, Looney RJ, Anderson CL, Abraham GN, Leddy JP. Human Fc gamma receptors: stable inter-donor variation in quantitative expression on platelets correlates with functional responses.. J Immunol. 1987; 138(9):2869-73. (Clone-specific: Flow cytometry). View Reference
      8. Sardjono, CT, Wines B, Powel M, Hogarth M. Epitope Mapping of Fc gamma RIIa Monoclonal Antibodies. Indonesian Journal of Biotechnology. 2008; 13:1030-1037. (Clone-specific: Flow cytometry).
      9. Su K, Yang H, Li X, et al. Expression profile of FcgammaRIIb on leukocytes and its dysregulation in systemic lupus erythematosus. J Immunol. 2007; 178(5):3272-3280. (Clone-specific). View Reference
      10. Van Den Herik Oudijk IE, Westerdaal NAC, De Haas M, et al. Binding heterogeneity within the CD32 panel of mAb. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:832-834.
      11. Van de Winkel JGJ, Anderson CL. CD32 cluster workshop report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:823-826.
      View All (11) View Less
      771047 Rev. 1

      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.