Multiparameter flow cytometric analysis of CD32 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Plot) or BD Horizon PE-CF594 Mouse Anti-Human CD32 antibody (Cat. No. 565647; 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.
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Multiparameter flow cytometric analysis of CD32 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Plot) or BD Horizon PE-CF594 Mouse Anti-Human CD32 antibody (Cat. No. 565647; 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
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_2739323
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™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- CF™ is a trademark of Biotium, Inc.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE-CF594 Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562305
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565647 Rev. 2
Antibody Details
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.
This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
565647 Rev. 2
Format Details
PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-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.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
565647 Rev.2
Citations & References
Development References (9)
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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
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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
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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
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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.
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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
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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.
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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
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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
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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.
565647 Rev. 2
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.
