Recombinant human anti-FcGR3A (also known as CD16A) biotinylated monoclonal antibody, which recognizes the human FcGR3A protein. This antibody has been tested for specific binding to purified human FcGR3A (BPS Bioscience #79013) in an ELISA binding assay with colorimetric detection. It does not cross-react with purified human FcGR2B (also known as CD16B).
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Synonyms
Low affinity immunoglobulin gamma Fc region receptor III-A, CD16a antigen, Fc-gamma RIII-alpha, FcR-10, IgG Fc receptor III-2, CD_antigen: CD16a, FcγRIIIA, FCGRIIIA
Protein A affinity chromatography from HEK293 supernatants.
Format
Aqueous buffer solution
Formulation
8 mM Phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
MW
Heavy Chain: 50 kDa; Light Chain: 25 kDa
UniProt #
P08637
Background
Fc Gamma Receptor IIIa (FcGR3A) is a low/intermediate affinity receptor for polyvalent immune-complexed IgG. It is involved in phagocytosis, antibody-dependent cytotoxicity, and clearance of immune complexes. FcGR3A plays a role in the activation of natural killer (NK) cells. Clinically, it serves as a marker of certain immune cells such as neutrophils. CD16 expression has been observed at the surface of T cells in patients with chronic viral infections (such as COVID-19). It is considered a potential therapeutic target to potentiate the efficacy of therapeutic antibodies used to treat solid tumors, or as direct target in hematopoietic cancers.
Storage/Stability
At least 12 months at -80°C.
Assay Conditions
Experimental design and assay protocol for measuring anti-FcGR3A specific binding to human FcGR3A in an ELISA assay:
Purified human FcGR3A (BPS Bioscience #79013) and purified human FcGR3B (BPS Bioscience #79016) were thawed on ice and coated onto a clear 96-well plate overnight at 4°C (4 µg/ml in PBS, 50 µl per well). **“No Coat” controls were included by coating PBS only to determine background levels**
The next day, the plate was washed three times with 1x Immuno Buffer 1 (BPS Bioscience #79311). Plate was tapped upside down on absorbent pads to remove excess liquid.
Wells were then blocked with 100 µl of Blocking Buffer 2 (BPS Bioscience #79728) for 1 hour at room temperature with slow shaking.
Serial dilutions of purified human biotin-labeled anti-FcGR3 were prepared in Blocking Buffer 2. **Titration from 0 nM to 300 nM, using 50 µl per well**
50 µl of the biotinylated antibody dilutions were added to the wells and incubated for 1 hour at room temperature with slow shaking. **Buffer only was added to wells designated as “BLANK”**
Wells were washed as in step 2.
Streptavidin-HRP (BPS Bioscience #79742) was diluted 1/1000 in Blocking Buffer 2. 50 µl was added to each well and incubated for 30-60 minutes at room temperature with slow shaking.
Wells were washed as in step 2.
100 µl of Colorimetric HRP Substrate (BPS Bioscience #79651) was added to all wells. Upon color development the reaction was quickly quenched with 100 µl of 1N HCl.
Absorbance was read at 450 nm. The “blank” value was subtracted from all other measurements.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Useful for studying the binding of FcGR3A in ELISA and in cellular assays.
Shipping Temperature
-80°C
Cross Reactivity
This antibody recognizes human FcGR3A/CD16A. It does not recognize human FcGR3B (CD16B). It has not been tested with other species
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