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RB780 Mouse Anti-Human FcγRIIA (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
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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. 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.
  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. An isotype control should be used at the same concentration as the antibody of interest.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. 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.
755694 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.

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Format Details
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RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
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RB780
Blue 488 nm
498 nm
781 nm
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Citations & References
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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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.