Anti-Hu CD16 Purified Low Endotoxin

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.

Product Specifications

Certification

RUO

Reactivity

Human

Immunogen

Human granulocytes

Target Antigen

CD16

Clone

MEM-154

Applications

FC (QC tested), IP, WB, FUNC

Concentration

1 mg/mL

Format

Purified Low Endotoxin

Buffer

Phosphate buffered saline (PBS), pH 7.4

References & Citations

*Gasdaska JR, Sherwood S, Regan JT, Dickey LF: An afucosylated anti-CD20 monoclonal antibody with greater antibody-dependent cellular cytotoxicity and B-cell depletion and lower complement-dependent cytotoxicity than rituximab. Mol Immunol. 2012 Mar;50 (3) :134-41, URL: https://pubmed.ncbi.nlm.nih.gov/22305040/, *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., URL: https://www.ncbi.nlm.nih.gov/pubmed/29780382 , *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., URL: https://www.ncbi.nlm.nih.gov/pubmed/8759741?dopt=A, *Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995) ., URL: , *de Haas M, Koene HR, Kleijer M, de Vries E, Simsek S, van Tol MJ, Roos D, von dem Borne AE: A triallelic Fc gamma receptor type IIIA polymorphism influences the binding of human IgG by NK cell Fc gamma RIIIa. J Immunol. 1996 Apr 15;156 (8) :3948-55., URL: https://www.ncbi.nlm.nih.gov/pubmed/8609432?dopt=A, *Koene HR, Kleijer M, Algra J, Roos D, von dem Borne AE, de Haas M: Fc gammaRIIIa-158V/F polymorphism influences the binding of IgG by natural killer cell Fc gammaRIIIa, independently of the Fc gammaRIIIa-48L/R/H phenotype. Blood. 1997 Aug 1;90 (3) :1109-14., URL: http://www.ncbi.nlm.nih.gov/pubmed/9242542 , *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., URL: https://www.sciencedirect.com/science/article/abs/pii/S0022175905002693?via%3Dihub

Other 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., URL: https://pubmed.ncbi.nlm.nih.gov/7836402/, *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., URL: https://pubmed.ncbi.nlm.nih.gov/9378982/, *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., URL: https://pubmed.ncbi.nlm.nih.gov/12626560/

Storage Conditions

Store at 2-8°C. Do not freeze.

Specificity

The antibody MEM-154 reacts with an extracellular epitope on CD16 antigen that is residing in proximity to FG loop (probably BC or C'E loop) . CD16 is a low affinity receptor for aggregated IgG (FcgammaRIII antigen) . The antibody MEM-154 reacts with CD16+ granulocytes, and it can be used for mapping CD16-158V/F polymorphism on NK cells, as it requires presence of V at amino acid 158.

Applications Notes

Functional application: The antibody MEM-154 blocks binding of human IgG to FcγRIII. Flow cytometry: Recommended dilution: 5-10 μg/ml; positive control: PBL (peripheral blood lymphocytes) . The antibody MEM-154 does not react with CD16a present on NK cells in many subjects. Western blotting: Non-reducing conditions.