Recombinant Human Fc gamma RIIIA/CD16a His Avi Protein, CF

When Human IgG is immobilized at 2 μg/mL, 100 μL/well, it binds Biotinylated Recombinant Human Fc gamma RIIIA/CD16a His-tag Avi-tag (AVI10468) with an ED50 of 4-40 ng/mL.

2 μg/lane of Biotinylated Recombinant Human Fc gamma RIIIA/CD16a His-tag Avi-tag (Catalog # AVI10468) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at kDa.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Fc gamma RIIIA/CD16a His Avi Protein, CF Summary

• Additional Information: Biotinylated Isoform V176
• Details of Functionality: The biotin to protein ratio is greater than 0.7 as determined by the HABA assay. Measured by its binding ability in a functional ELISA. When Human IgG is immobilized at 2 μg/mL, 100 μL/well, it binds Biotinylated Recombinant Human Fc gamma RIIIA/CD16a His-tag Avi-tag (Catalog # AVI10468) with an ED₅₀ of 4-40 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived human Fc gamma RIIIA/CD16a protein
  

  Human Fc gamma RIIIA/CD16a (Gly17-Gln208) Accession # AAH17865.1	6-His tag	Avi-tag
  N-terminus		C-terminus

• Accession #: AAH17865.1
• N-terminal Sequence: Gly17
• Structure / Form: Biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
  • Bioactivity2
• Theoretical MW: 24 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: 48-57 kDa, under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human Fc gamma RIIIA/CD16a His Avi Protein, CF

• 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/CD16a 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 (1-3). In humans, it is expressed as a 50-70 kDa transmembrane activating receptor on NK cells, T cells, monocytes, and macrophages (2). It is closely related to the GPI-linked Fc gamma RIIIB which is expressed on human neutrophils and eosinophils (1, 3). These two proteins share 97% amino acid (aa) identity within their extracellular domains (ECD) (4). The ECD of Fc gamma RIIIA also 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, respectively. Mature human Fc gamma RIIIA consists of a 192 aa ECD with two C2-type Ig-like domains, a 21 aa transmembrane segment, and a 25 aa cytoplasmic domain. In humans, a single nucleotide polymorphism (T230A) creates high binding (176V) and low binding (176F) forms that may influence susceptibility to autoimmune diseases or response to therapeutic IgG antibodies (5, 6). Fc gamma RIIIA surface expression requires interaction with an accessory chain, either the common gamma -chain or CD3 zeta (7, 8). Glycosylation patterns, electrophoretic mobility, and binding affinity appear to differ between NK cell and monocyte Fc gamma RIIIA (9). Shed forms of both Fc gamma RIIIA and Fc gamma RIIIB can be generated by proteolytic cleavage and retain binding activity (10-13). Shedding from monocytes and macrophages can be triggered by cell activation or phagocytosis (13). Soluble Fc gamma RIII circulates in normal plasma and is elevated in rheumatoid arthritis and in coronary artery diseases (11, 12). Our Avi-tag Biotinylated human Fc gamma RIIIA features biotinylation at a single site contained within the Avi-tag, a unique 15 aa peptide.  Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

1. Nagelkerke, S.Q. and T.W. Kuijpers (2015) Front. Immunol. 5:674.
2. Nimmerjahn, F. and J.V. Ravetch (2006) Immunity 24:19.
3. Ravetch, J.V. and B. Perussia (1989) J. Exp. Med. 170:481.
4. Scallon, B.J. et al. (1989) Proc. Natl. Acad. Sci. USA 86:5079.
5. Wu, J. et al. (1997) J. Clin. Invest. 100:1059.
6. Dall’Ozzo, S. et al. (2004) Cancer Res. 64:4664.
7. Kim, M.-K. et al. (2003) Blood 101:4479.
8. Lanier, L.L. et al. (1989) Nature 342:803.
9. Edberg, J.C. and R.P. Kimberley (1997) J. Immunol. 159:3849.    
10. Li, P. et al. (2007) J. Biol. Chem. 282:6210.
11. Masuda, M. et al. (2003) J. Rheumatol. 30:1911.
12. Masuda, M. et al. (2006) Atherosclerosis 188:377.
13. Webster, N.L. et al. (2006) J. Leukoc. Biol. 79:294.

FAQs for Fc gamma RIIIA/CD16a (AVI10468). (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()