Anti-Hu CD16 FITC

Anti-Hu CD16 FITC
Regulatory status
RUO
Antigen
CD16
Clone
MEM-154
Format
FITC
Reactivity
Human
Application
Excitation laser
blue (488 nm)
Variant
100 tests
1F-214-T100
In stock
198.00 USD

25 tests
1F-214-T025
Delivery 1 week
99.00 USD
Variant
100 tests
1F-214-T100
In stock
198.00 USD

25 tests
1F-214-T025
Delivery 1 week
99.00 USD
Product details
Description
Images
References
SDS download
Isotype
Mouse IgG1
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.
Workshop
HLDA V: WS Code M MA068; HLDA V: WS Code NK NK51
Application
Application details
Flow cytometry: The reagent is designed for analysis of human blood cells using 20 μl reagent / 100 μl of whole blood or 106 cells in a suspension. The content of a vial (2 ml) is sufficient for 100 tests.
Reactivity
Human
Immunogen
Human granulocytes
Preparation
Purified antibody is conjugated with fluorescein isothiocyanate (FITC) 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.
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.
1F-214_FC_CDMaps-Profil značení
Anti-human CD16 FITC antibody (clone MEM-154) 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 FITC antibody (clone MEM-154) was used in concentration 6.7 µg/ml in stained blood sample (2 x 106 cells).
Donor of tested blood sample had 158F polymorphism on CD16a.
1F-214_FC_CDMaps-Histogram
Reactivity of Anti-human CD16 FITC antibody (clone MEM-154) 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 FITC antibody (clone MEM-154, 6.7 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.
Donor of tested blood sample had 158F polymorphism on CD16a.

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:

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
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
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
Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995).
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.
PubMed
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.
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
Variant
100 tests
1F-214-T100
In stock
198.00 USD

25 tests
1F-214-T025
Delivery 1 week
99.00 USD
Variant
100 tests
1F-214-T100
In stock
198.00 USD

25 tests
1F-214-T025
Delivery 1 week
99.00 USD

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