Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein New

Name: Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein
Brand: R&D Systems
SKU: AFG4325

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AFG4325

Product Datasheet / COA / SDS

His-tag

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Setting the standard in quality research reagents

Key Product Details

Source:	NS0
Accession #:	AAH17865.1
Structure / Form:	Labeled with Alexa Fluor® 488 via amines Excitation Wavelength: 488 nm Emission Wavelength: 515–545 nm
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

Product Specifications for Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

Source	Mouse myeloma cell line, NS0-derived human Fc gamma RIIIA/CD16a protein Gly17-Gln208, with a C-terminal 6-His tag
Purity	>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Gly17
Predicted Molecular Mass	22.6 kDa
SDS-PAGE	40-50 kDa, under reducing conditions.
Activity	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.

Scientific Data Examples for Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

	Flow cytometry analysis for Recombinant Human Fc gamma RIIIA/CD16a Alexa Fluor® 488 staining on Human Fc gamma RIIIA/CD16a Antibody conjugated beads. 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).
	Recombinant Human Fc gamma RIIIA/CD16a Alexa Fluor® 488 Protein SDS-PAGE. 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.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

AFG4325

Formulation	Supplied as a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Reconstitution Calculator

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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).

References

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

Product Specific Notices for Recombinant Human Fc gamma RIIIA Alexa Fluor® 488 Protein

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.

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