Anti-Hu CD16 PE-Cy™5

Anti-Hu CD16 PE-Cy™5
Regulatory status
RUO
Antigen
CD16
Clone
3G8
Format
PE-Cy™5
Reactivity
Non-human primates, Human
Application
Excitation laser
blue (488 nm)
Variant
100 tests
T8-646-T100
In stock
242.00 USD
Variant
100 tests
T8-646-T100
In stock
242.00 USD
Product details
Description
Images
References
SDS download
Isotype
Mouse IgG1 kappa
Specificity
The mouse monoclonal antibody 3G8 recognizes an extracellular epitope of CD16, a low affinity receptor for aggregated IgG (FcgammaRIII antigen). CD16 exists in two different isoforms: CD16a (FcgammaRIIIA; 50-65 kDa; expressed on NK-cells, monocytes and macrophages) and CD16b (FcgammaRIIIB; 48 kDa; mainly expressed on neutrophils). Regarding CD16-158V/F polymorphism, the antibody 3G8 detects both 158V and 158F allotype on natural NK cells.
Workshop
HLDA V: WS Code NK80
Application
Application details
Flow cytometry: The reagent is designed for analysis of human blood cells using 4 μl reagent / 100 μl of whole blood or 106 cells in a suspension. The content of a vial (0.4 ml) is sufficient for 100 tests.
Reactivity
Non-human primates, Human
Immunogen
Human neutrophils
Preparation
Purified antibody is conjugated with activated tandem dye of R-phycoerythrin-cyanine 5 (PE-Cy™5) under optimum conditions and unconjugated antibody and free fluorochrome are removed by size-exclusion chromatography.
Formulation
Stabilizing phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Storage and handling
Store at 2-8°C. Protect from prolonged exposure to light. Do not freeze.
Exbio licence note
The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.
Licence note
Cy™ and CyDye™ are registered trademarks of Cytiva.
Other names
FcgammaRIII, IGFR3, FCRIII
Antigen description
CD16 (FcgammaRIII) is a 50-65 kDa glycoprotein serving as a low affinity IgG receptor. Human FcgammaRIII is expressed in two forms – FcgammaRIII-A and -B. FcgammaRIII-A is a transmembrane protein of monocytes, macrophages, NK cells and a subset of T cells. It is associated with FcepsilonRI-gamma subunit and is responsible for antibody-dependent NK cell cytotoxicity. Mast cell FcgammaRIII-A is associated, moreover, with FcepsilonRI-beta subunit. Besides IgG, FcgammaRIII-A can be triggered also by oligomeric IgE. FcgammaRIII-B is a GPI-linked monomeric receptor expressed on neutrophils and is involved in their activation and induction of a proadhesive phenotype.
T8-646_FC_CDMaps_Profil značení
Anti-human CD16 PE-Cy™5 antibody (clone 3G8) works in flow cytometry application.

Analysis of the antibody staining profile was performed on blood leukocytes isolated from peripheral whole blood by bulk erythrocyte lysis using 10× diluted EXCELLYSE Live (ED7068).
Mouse monoclonal anti-human CD16 PE-Cy™5 antibody (clone 3G8) was used in concentration 9 µg/ml in stained blood sample (2 x 106 cells).
T8-646_FC_CDMaps_Fot-plot
Staining pattern of CD16 PE-Cy™5 antibody (clone 3G8) in dot-plot fluorescence visualization (lymphocyte gate)

Analysis of the antibody staining profile was performed on blood leukocytes isolated from peripheral whole blood by bulk erythrocyte lysis using 10× diluted EXCELLYSE Live (ED7068).
Mouse monoclonal anti-human CD16 PE-Cy™5 antibody (clone 3G8) was used in concentration 9 µg/ml in stained stained blood sample (2 x 106 cells) and Mouse monoclonal anti-human CD3 FITC antibody (clone UCHT1) in concentration 5 µg/ml, respectively.
T8-646_FC_CDMaps_Dot-plot2
Staining pattern of CD16 PE-Cy™5 antibody (clone 3G8) in dot-plot fluorescence visualization (mononuclear gate)

Analysis of the antibody staining profile was performed on blood leukocytes isolated from peripheral whole blood by bulk erythrocyte lysis using 10× diluted EXCELLYSE Live (ED7068).
Mouse monoclonal anti-human CD16 PE-Cy™5 antibody (clone 3G8) was used in concentration 9 µg/ml in stained stained blood sample (2 x 106 cells) and Mouse monoclonal anti-human CD14 APC-Cy™7 antibody (clone MEM-15) in concentration 4 µg/ml, respectively.
T8-646_FC_CDMaps_Histogram
Reactivity of Anti-human CD16 PE-Cy™5 antibody (clone 3G8) on human peripheral leukocytes

Analysis of the antibody staining profile was performed on blood leukocytes isolated from peripheral whole blood by bulk erythrocyte lysis using 10× diluted EXCELLYSE Live (ED7068).
Suspension of blood leukocytes (2 x 106 cells) was added to the mixture of CD16 PE-Cy™5 antibody (clone 3G8, 9 µg/ml in stained blood sample), backbone antibody conjugates and Monocyte Blocking Buffer (#ED7747), vortexed and incubated for 20 min. Stained sample was fixed with 2 ml of 10× diluted EXCELLYSE Easy solution (#ED7066) for 10 min. Finally, samples were centrifuged (670 g, 5 min.), supernatant removed and the cell pellet was resuspended in 200 µl of PBS for acquisition.
T8-646_FC_Dot-plot
Flow cytometry multicolor surface staining of human lymphocytes stained using anti-human CD16 (3G8) PE-DyLight® 594 antibody (4 μl reagent / 100 μl of peripheral whole blood) and anti-human CD3 (UCHT1) APC antibody (10 μl reagent / 100 μl of peripheral whole blood).
T8-646_FC_Histogram
Separation of human CD16 positive CD3 negative NK cells (red-filled) from CD16 negative CD3 positive T cells (black-dashed) in flow cytometry analysis (surface staining) of human peripheral whole blood stained using anti-human CD16 (3G8) PE-Cy™5 antibody (4 μl reagent / 100 μl of peripheral whole blood).
T8-646_FC_Profil Značení
Flow cytometry surface staining pattern of human peripheral whole blood stained using anti-human CD16 (3G8) PE-Cy™5 antibody (4 μl reagent / 100 μl of peripheral whole blood).

General references:

Gessner JE, Grussenmeyer T, Kolanus W, Schmidt RE: The human low affinity immunoglobulin G Fc receptor III-A and III-B genes. Molecular characterization of the promoter regions. J Biol Chem. 1995 Jan 20;270(3):1350-61.
PubMed
Kocher M, Siegel ME, Edberg JC, Kimberly RP: Cross-linking of Fc gamma receptor IIa and Fc gamma receptor IIIb induces different proadhesive phenotypes on human neutrophils. J Immunol. 1997 Oct 15;159(8):3940-8.
PubMed
Arase N, Arase H, Hirano S, Yokosuka T, Sakurai D, Saito T: IgE-mediated activation of NK cells through Fc gamma RIII. J Immunol. 2003 Mar 15;170(6):3054-8.
PubMed

Product specific references:

Zhu X, Hamann KJ, Muñoz NM, Rubio N, Mayer D, Herrnreiter A, Leff AR: Intracellular expression of Fc gamma RIII (CD16) and its mobilization by chemoattractants in human eosinophils. J Immunol. 1998 Sep 1;161(5):2574-9.
PubMed
Jeraiby M, Sidi Yahya K, Depince-Berger AE, Lambert C: Microbicidal activity measured by flow cytometry: Optimization and standardization for detection of primary and functional deficiencies. J Immunol Methods. 2016 Sep 29. pii: S0022-1759(16)30220-4.
PubMed
Ohradanova-Repic A, Machacek C, Charvet C, Lager F, Le Roux D, Platzer R, Leksa V, Mitulovic G, Burkard TR, Zlabinger GJ, Fischer MB, Feuillet V, Renault G, Blüml S, Benko M, Suchanek M, Huppa JB, Matsuyama T, Cavaco-Paulo A, Bismuth G, Stockinger H: Extracellular Purine Metabolism Is the Switchboard of Immunosuppressive Macrophages and a Novel Target to Treat Diseases With Macrophage Imbalances. Front Immunol. 2018 Apr 27;9:852.
PubMed
Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989).
Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995).
Zhu X, Hamann KJ, Muñoz NM, Rubio N, Mayer D, Herrnreiter A, Leff AR: Intracellular expression of Fc gamma RIII (CD16) and its mobilization by chemoattractants in human eosinophils. J Immunol. 1998 Sep 1;161(5):2574-9.
PubMed
Metes D, Ernst LK, Chambers WH, Sulica A, Herberman RB, Morel PA: Expression of functional CD32 molecules on human NK cells is determined by an allelic polymorphism of the FcgammaRIIC gene. Blood. 1998 Apr 1;91(7):2369-80.
PubMed
Wijngaarden S, van Roon JA, van de Winkel JG, Bijlsma JW, Lafeber FP: Down-regulation of activating Fcgamma receptors on monocytes of patients with rheumatoid arthritis upon methotrexate treatment. Rheumatology (Oxford). 2005 Jun;44(6):729-34.
PubMed
Komano Y, Nanki T, Hayashida K, Taniguchi K, Miyasaka N: Identification of a human peripheral blood monocyte subset that differentiates into osteoclasts. Arthritis Res Ther. 2006;8(5):R152.
PubMed
Choi EI, Wang R, Peterson L, Letvin NL, Reimann KA: Use of an anti-CD16 antibody for in vivo depletion of natural killer cells in rhesus macaques. Immunology. 2008 Jun;124(2):215-22. Epub 2008 Jan 12.
PubMed
Congy-Jolivet N, Bolzec A, Ternant D, Ohresser M, Watier H, Thibault G: Fc gamma RIIIa expression is not increased on natural killer cells expressing the Fc gamma RIIIa-158V allotype. Cancer Res. 2008 Feb 15;68(4):976-80.
PubMed
Burt BM, Plitas G, Zhao Z, Bamboat ZM, Nguyen HM, Dupont B, DeMatteo RP: The lytic potential of human liver NK cells is restricted by their limited expression of inhibitory killer Ig-like receptors. J Immunol. 2009 Aug 1;183(3):1789-96.
PubMed
Böttcher S, Ritgen M, Brüggemann M, Raff T, Lüschen S, Humpe A, Kneba M, Pott C: Flow cytometric assay for determination of FcγRIIIA-158 V/F polymorphism. J Immunol Methods 2005, 306, 128-136.
PubMed
Hoshino S, Oshimi K, Teramura M, Mizoguchi H: Activation via the CD3 and CD16 pathway mediates interleukin-2-dependent autocrine proliferation of granular lymphocytes in patients with granular lymphocyte proliferative disorders. Blood 1991 Dec 15;78(12):3232-40.
PubMed
Tamm A, Schmidt RE: The binding epitopes of human CD16 (Fc gamma RIII) monoclonal antibodies. Implications for ligand binding. J Immunol. 1996 Aug 15;157(4):1576-81.
PubMed
Holl V, Hemmerter S, Burrer R, Schmidt S, Bohbot A, Aubertin AM, Moog C: Involvement of Fc gamma RI (CD64) in the mechanism of HIV-1 inhibition by polyclonal IgG purified from infected patients in cultured monocyte-derived macrophages. J Immunol. 2004 Nov 15;173(10):6274-83.
PubMed
Hober D, Chehadeh W, Weill J, Hober C, Vantyghem MC, Gronnier P, Wattré P: Circulating and cell-bound antibodies increase coxsackievirus B4-induced production of IFN-alpha by peripheral blood mononuclear cells from patients with type 1 diabetes. J Gen Virol. 2002 Sep;83(Pt 9):2169-2176.
PubMed
Quardokus EM, Saunders DC, McDonough E, Hickey JW, Werlein C, Surrette C, Rajbhandari P, Martinez Casals A, Tian H, Lowery L, Neumann EK, Björklund F, Neelakantan TV, Croteau J, Wiblin AE, Fisher J, Livengood AJ, Dowell KG, Silverstein JC, Spraggins JM, Pryhuber GS, Deutsch G, Ginty F, Nolan GP, Melov S, Jonigk D, Caldwell MA, Vlachos IS, Mueller W, Gehlenborg N, Stockwell BR, Lundberg E, Snyder MP, Germain RN, Camarillo JM, Kelleher NL, Börner K, Radtke AJ: Organ mapping antibody panels: a community resource for standardized multiplexed tissue imaging. Nat Methods 2023 Aug;20(8):1174-1178. doi: 10.1038/s41592-023-01846-7.
PubMed
Variant
100 tests
T8-646-T100
In stock
242.00 USD
Variant
100 tests
T8-646-T100
In stock
242.00 USD

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