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BV421 Mouse Anti-Human CD32
BV421 Mouse Anti-Human CD32

Multiparameter flow cytometric analysis of CD32 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD32 antibody (Cat. No. 564838; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).  Two-parameter flow cytometric contour plots showing the correlated expression of CD32 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Multiparameter flow cytometric analysis of CD32 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or BD Horizon BV421 Mouse Anti-Human CD32 antibody (Cat. No. 564838; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202).  Two-parameter flow cytometric contour plots showing the correlated expression of CD32 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
CD32a,FcγRIIa, FCGR2A; CD32b, FcγRIIb, FCGR2B; CD32c, FcγRIIc, FCGR2C
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG2b, κ
K562 or FcγRII+ L cell lines
Flow cytometry (Routinely Tested)
5 µl
V MA128
AB_2738976
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

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).

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. An isotype control should be used at the same concentration as the antibody of interest.
564838 Rev. 1
Antibody Details
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FLI8.26

The FLI8.26 monoclonal antibody specifically recognizes CD32 which is also known as Fcγ receptor type II (FcγRII). CD32 is a type I transmembrane glycoprotein that serves as a low affinity receptor for aggregated IgG. The FLI8.26 antibody recognizes a, b, and c forms of CD32 that are encoded by separate genes: CD32a/FcγRIIa (FCGR2A), CD32b/FcγRIIb (FCGR2B), and CD32c/FcγRIIc (FCGR2C). These forms are differentially expressed on monocytes, granulocytes, T cells, B cells, NK cells, or platelets. These receptors play various roles in mediating and regulating inflammation and immunity including phagocytosis, cytotoxicity, degranulation, or platelet activation.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564838 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564838 Rev.1
Citations & References
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Development References (9)

  1. Båve U, Magnusson M, Eloranta ML, Perers A, Alm GV, Rönnblom L. Fc gamma RIIa is expressed on natural IFN-alpha-producing cells (plasmacytoid dendritic cells) and is required for the IFN-alpha production induced by apoptotic cells combined with lupus IgG.. J Immunol. 2003; 171(6):3296-302. (Clone-specific: Flow cytometry). View Reference
  2. Hogarth PM, Anania JC, Wines BD. The FcγR of humans and non-human primates and their interaction with IgG: implications for induction of inflammation, resistance to infection and the use of therapeutic monoclonal antibodies.. Curr Top Microbiol Immunol. 2014; 382:321-52. (Biology). View Reference
  3. Ierino FL, Hulett MD, McKenzie IF, Hogarth PM. Mapping epitopes of human Fc gamma RII (CDw32) with monoclonal antibodies and recombinant receptors. J Immunol. 1993; 150(5):1794-1803. (Immunogen). View Reference
  4. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  5. Meknache N, Jönsson F, Laurent J, Guinnepain MT, Daëron M. Human basophils express the glycosylphosphatidylinositol-anchored low-affinity IgG receptor FcgammaRIIIB (CD16B).. J Immunol. 2009; 182(4):2542-50. (Clone-specific: Flow cytometry). View Reference
  6. 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.
  7. Stuart SG, Simister NE, Clarkson SB, Kacinski BM, Shapiro M, Mellman I. Human IgG Fc receptor (hFcRII; CD32) exists as multiple isoforms in macrophages, lymphocytes and IgG-transporting placental epithelium. EMBO J. 1989; 8(12):3657-3666. (Biology). View Reference
  8. Tomiyama Y, Kunicki TJ, Zipf TF, Ford SB, Aster RH. Response of human platelets to activating monoclonal antibodies: importance of Fc gamma RII (CD32) phenotype and level of expression. Blood. 1992; 80(9):2261-2268. (Biology). View Reference
  9. 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.
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564838 Rev. 1

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