Flow cytometric analysis of CD16/CD32 expressed on mouse splenic lymphocytes. (Left Panel) Splenocytes from BALB/c mice were stained with either the PE-Cy™7 Rat Anti-Mouse CD16/CD32 antibody (Cat. No. 560829; solid line histogram) or a PE-Cy™7 Rat IgG2b, κ Isotype Control (Cat. No. 552849; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable splenic lymphocytes. Splenocytes from BALB/c mice were stained with either a PE-Cy™7 Rat IgG2b, κ isotype control (Cat. No. 552849; Middle Panel) or with PE-Cy™7 Rat Anti-Mouse CD16/CD32 antibody (Cat. No. 560829; Right Panel) in conjunction with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066). Two color fluorescent dot plots were derived from gated events with the forward and side light-scatter characteristics of viable splenic lymphocytes. Flow cytometry was performed on a BD LSR™ II Flow Cytometer System.
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Flow cytometric analysis of CD16/CD32 expressed on mouse splenic lymphocytes. (Left Panel) Splenocytes from BALB/c mice were stained with either the PE-Cy™7 Rat Anti-Mouse CD16/CD32 antibody (Cat. No. 560829; solid line histogram) or a PE-Cy™7 Rat IgG2b, κ Isotype Control (Cat. No. 552849; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable splenic lymphocytes. Splenocytes from BALB/c mice were stained with either a PE-Cy™7 Rat IgG2b, κ isotype control (Cat. No. 552849; Middle Panel) or with PE-Cy™7 Rat Anti-Mouse CD16/CD32 antibody (Cat. No. 560829; Right Panel) in conjunction with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066). Two color fluorescent dot plots were derived from gated events with the forward and side light-scatter characteristics of viable splenic lymphocytes. Flow cytometry was performed on a BD LSR™ II Flow Cytometer System.
Product Details
BD Pharmingen™
FcγRIII/FcγRII; Fcgr3/Fcgr2
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Mouse BALB/c Macrophage J774
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_10563207
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. 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 PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- 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.
- Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
PE-Cy™7 Rat IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
552849
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APC Hamster Anti-Mouse CD3e RUO
Size
0.1 mg
Cat No.
553066
560829 Rev. 1
Antibody Details
2.4G2
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.
560829 Rev. 1
Format Details
PE-Cy7
PE-Cy7 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, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-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 Red (627–640-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-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560829 Rev.1
Citations & References
Development References (19)
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Jensen WA, Marschner S, Ott VL, Cambier JC. FcgammaRIIB-mediated inhibition of T-cell receptor signal transduction involves the phosphorylation of SH2-containing inositol 5-phosphatase (SHIP), dephosphorylation of the linker of activated T-cells (LAT) and inhibition of calcium mobilization. Biochem Soc Trans. 2001; 29(6):840-846. (Biology). View Reference
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Katz HR, Arm JP, Benson AC, Austen KF. Maturation-related changes in the expression of Fc gamma RII and Fc gamma RIII on mouse mast cells derived in vitro and in vivo. J Immunol. 1990; 145(10):3412-3417. (Biology). 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. (Biology). View Reference
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Lewis VA, Koch T, Plutner H, Mellman I. A complementary DNA clone for a macrophage-lymphocyte Fc receptor. Nature. 1986; 324(6095):372-375. (Biology). View Reference
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Perussia B, Tutt MM, Qiu WQ, et al. Murine natural killer cells express functional Fc gamma receptor II encoded by the Fc gamma R alpha gene. J Exp Med. 1989; 170(1):73-86. (Biology). View Reference
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Rodewald HR, Awad K, Moingeon P, et al. Fc gamma RII/III and CD2 expression mark distinct subpopulations of immature CD4-CD8- murine thymocytes: in vivo developmental kinetics and T cell receptor beta chain rearrangement status. J Exp Med. 1993; 177(4):1079-1092. (Biology). View Reference
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Rodewald HR, Moingeon P, Lucich JL, Dosiou C, Lopez P, Reinherz EL. A population of early fetal thymocytes expressing Fc gamma RII/III contains precursors of T lymphocytes and natural killer cells. Cell. 1992; 69(1):139-150. (Biology). View Reference
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Takezawa R, Watanabe Y, Akaike T. Direct evidence of macrophage differentiation from bone marrow cells in the liver: a possible origin of Kupffer cells. J Biochem (Tokyo). 1995; 118(6):1175-1183. (Biology). 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. (Biology). View Reference
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560829 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.
