Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

Streptavidin coated beads conjugated to Human Fc gamma RIII (CD16) Biotinylated Antibody (FAB2546B) were stained with the indicated concentrations of Recombinant Human Fc gamma RIIIA/CD16a Alexa Fluor® 488 (Catalog # AFG4325).

2 μg/lane of Recombinant Human Fc gamma RIIIA/CD16a Alexa Fluor® 488 Protein (Catalog # AFG4325) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 40-50 kDa.

• Applications: Bioactivity

Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein Summary

• Additional Information: His-tag
• Details of Functionality: Measured by flow cytometry for its ability to bind anti-human Fc gamma RIIIA/CD16a Antibody conjugated beads. The concentration of Recombinant Human Fc gamma RIIIA/CD16a Alexa Fluor® 488 (Catalog# AFG4325) that produces 50% of the binding response is 15.0-150 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived human Fc gamma RIIIA/CD16a protein
  Gly17-Gln208, with a C-terminal 6-His tag
• Accession #: AAH17865.1
• N-terminal Sequence: Gly17
• Structure / Form: Labeled with Alexa Fluor® 488 via amines
  Excitation Wavelength: 488 nm
  Emission Wavelength: 515–545 nm
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 22.6 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 40-50 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Notes

This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

• CD16a antigen
• CD16a
• CD16FCRIIIA
• Fc fragment of IgG, low affinity IIIa, receptor (CD16a)
• Fc fragment of IgG, low affinity IIIa, receptor for (CD16)
• Fc gamma receptor III-A
• Fc gamma RIIIA
• FCG3
• Fc-gamma receptor III-2 (CD 16)
• Fc-gamma receptor IIIb (CD16)
• Fc-gamma RIII
• Fc-gamma RIIIa
• Fc-gamma RIII-alpha
• FCGR3
• FCGR3A
• FCGRIII
• FcgRIIIA
• FcR-10
• FCRIII
• FcRIIIa
• IGFR3FcgammaRIIIA
• immunoglobulin G Fc receptor III
• low affinity III, receptor for (CD16)
• low affinity immunoglobulin gamma Fc region receptor III-A
• neutrophil-specific antigen NA

Background

Fc gamma RIIIa is a low/intermediate affinity receptor for polyvalent immune-complexed IgG. It is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody-dependent cytotoxicity and clearance of immune complexes (1, 2). In humans, it is a 50-70 kDa type I transmembrane activating receptor expressed by NK cells, T cells, monocytes, and macrophages (1). Fc gamma RIIIb is highly related, sharing 97% amino acid (aa) identity within the extracellular domain (ECD), but is a GPI-linked receptor expressed on human neutrophils and eosinophils (1, 2). The ECD of Fc gamma RIIIa shares 63%, 61%, 65%, 59% and 58% aa identity with mouse Fc gamma RIV, rat Fc gamma RIIIa, feline CD16, bovine CD16 and porcine Fc gamma RIIIb paralogs, respectively. The Fc gamma RIIIa cDNA encodes 254 aa including a 16 aa signal sequence, 191 aa ECD with two C2-type Ig-like domains and five potential N-glycosylation sites, a 22 aa transmembrane (TM) sequence and a 25 aa cytoplasmic domain. In humans, a single nucleotide polymorphism creates high binding (176V) and low binding (176F) forms that, when homozygous, may influence susceptibility to autoimmune diseases or response to therapeutic IgG antibodies (3, 4). Catalog # 4325-FC is expressed as the 176V isoform of Fc gamma RIIIa. Fc gamma RIIIa surface expression requires interaction of an accessory chain, either the common gamma -chain or CD3 zeta (5, 6). Glycosylation patterns, electrophoretic mobility and binding affinity appear to differ between NK cell and monocyte Fc gamma RIIIa (7). The ECD of both Fc gamma RIIIa and b can be proteolytically cleaved and retain binding activity in soluble form (8-11). In monocytes and macrophages, activation and phagocytosis can trigger Fc gamma RIIIa release (11). Soluble Fc gamma RIII can be detected in normal plasma and is increased in rheumatoid arthritis and in coronary artery diseases (9, 10).

1. Nimmerjahn, F. and J.V. Ravetch (2006) Immunity 24:19. 
2. Ravetch, J.V. and B. Perussia (1989) J. Exp. Med. 170:481. 
3. Wu, J. et al. (1997) J. Clin. Invest. 100:1059. 
4. Dall’Ozzo, S. et al. (2004) Cancer Res. 64:4664. 
5. Kim, M.-K. et al. (2003) Blood 101:4479. 
6. Lanier, L.L. et al. (1989) Nature 342:803.
7. Edberg, J.C. and R.P. Kimberley (1997) J. Immunol. 159:3849.
8. Li, P. et al. (2007) J. Biol. Chem. 282:6210.
9. Masuda, M. et al. (2003) J. Rheumatol. 30:1911.
10. Masuda, M. et al. (2006) Atherosclerosis 188:377.
11. Webster, N.L. et al. (2006) J. Leukoc. Biol. 79:294.

FAQs for Fc gamma RIIIA/CD16a (AFG4325). (Showing 1 - 1 of 1 FAQ).

1. I am interested in developing a surrogate ADCC assay (FC gamma III a binding assay) for which, I would require a CD16 Recombinant Protein as an antigen and Anti-Human IgG (Fc specific) as a secondary antibody as I am planning to perform this assay in an ELISA format. With this brief description, could you please suggest me the right CD16 Recombinant Protein among the three listed products in your web site (www.novusbio.com/proteins/cd16). Appreciate if you can share a reference article, where the use of this protein is cited.
   • The right CD16 protein for you depends on what sort of functionality you require for your assay. H00002214-P01 is made by a Taiwanese company called Abnova and we distribute it for them. They do not test the activity of their recombinant proteins. The proteins are produced in a cell-free wheat germ system with an N-terminal 26kDa GST-tag. Due to this expression system, in our experience, the proteins do not always undergo the same folding and post-transnational modifications they might undergo in an endogenous cell. Because of this, we do not recommend this protein for use in functional assays and do not guarantee them for this use. They are mainly intended for use in Western Blots and ELISA as a positive control with their antibody. NBP1-98908 is produced in E. coli and contains a His-tag and it also not guaranteed for functionality but may be due to its E. coli expression system. H00002214-G01 is also an Abnova and is made in their cell-free wheat germ expression system with proprietary liposome technology. This is a recently developed system by them and is intended to help the proteins fold and maintain functionality. Which protein would be best for you depends on what you require for your assay. Here is a link to our antibodies to the Fc region of human IgG. The best antibody for you will depend on what you would like to use for detection.

Bioinformatics

• Uniprot:
  • AAH17865.1()