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Two-color flow cytometric analysis of CD16/32 expression on mouse splenocytes. Splenic leucocytes were stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either BD Horizon™ BUV737 Rat IgG2b, κ Isotype Control (Cat. No. 564295; Left Panel) or BD Horizon BUV737 Rat Anti-Mouse CD16/CD32 antibody (Cat. No. 565272; Right Panel). Two-color flow cytometric contour plots show the correlated expression patterns of CD16/CD32 (or Ig Isotype control staining) versus CD3e for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BUV737 Rat Anti-Mouse CD16/CD32
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 2.4G2 antibody specifically recognizes a common nonpolymorphic epitope on the extracellular domains of the mouse FcγIII (CD16) and FcγII (CD32) Receptors. It has also been reported to bind the FcγI receptor (CD64) via its Fc domain. 2.4G2 mAb blocks non-antigen-specific binding of immunoglobulins to the FcγIII and FcγII, and possibly FcγI, Receptors in vitro and in vivo. CD16 and/or CD32 are expressed on natural killer cells, monocytes, macrophages, dendritic cells (at low levels), Kupffer cells, granulocytes, mast cells, B lymphocytes, immature thymocytes, and some activated mature T lymphocytes.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (e.g., 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (e.g., CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (7)
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Araujo-Jorge T, Rivera MT, el Bouhdidi A, Daeron M, Carlier Y. An Fc gamma RII-, Fc gamma RIII-specific monoclonal antibody (2.4G2) decreases acute Trypanosoma cruzi infection in mice. Infect Immun. 1993; 61(11):4925-4928. (Clone-specific: Flow cytometry, Functional assay, In vivo exacerbation). View Reference
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Kurlander RJ, Ellison DM, Hall J. The blockade of Fc receptor-mediated clearance of immune complexes in vivo by a monoclonal antibody (2.4G2) directed against Fc receptors on murine leukocytes. J Immunol. 1984; 133(2):855-862. (Clone-specific: Functional assay, In vivo exacerbation, Radioimmunoassay). View Reference
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Latour S, Bonnerot C, Fridman WH, Daeron M. Induction of tumor necrosis factor-alpha production by mast cells via Fc gamma R. Role of the Fc gamma RIII gamma subunit. J Immunol. 1992; 149(6):2155-2162. (Clone-specific: Flow cytometry, Functional assay, Stimulation). View Reference
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Maeda K, Burton GF, Padgett DA, et al. Murine follicular dendritic cells and low affinity Fc receptors for IgE (Fc epsilon RII). J Immunol. 1992; 148(8):2340-2347. (Clone-specific: Immunohistochemistry). View Reference
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Mellman IS, Unkeless JC. Purificaton of a functional mouse Fc receptor through the use of a monoclonal antibody. J Exp Med. 1980; 152(4):1048-1069. (Clone-specific: Immunoaffinity chromatography, Immunoprecipitation, Radioimmunoassay). View Reference
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Titus JA, Finkelman FD, Stephany DA, Jones JF, Segal DM. Quantitative analysis of Fc gamma receptors on murine spleen cell populations by using dual parameter flow cytometry. J Immunol. 1984; 133(2):556-561. (Clone-specific: Flow cytometry). View Reference
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Unkeless JC. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979; 150(3):580-596. (Immunogen: Fluorescence microscopy, Immunofluorescence, Inhibition, Radioimmunoassay). View Reference
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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.