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Biotinylated Human Fc gamma RIIIA / CD16a (V176), Avi Tag (Avitag™)  pdf  pdf  pdf


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Synonym

FCGR3A,CD16A,FCG3,FCGR3,IGFR3

Source

MABSol® Biotinylated Human Fc gamma RIIIA / CD16a (V176)(CDA-H82E9) is expressed from human HEK293 cells. It contains AA Gly 17 - Gln 208 (Accession # P08637-1).

Predicted N-terminus: Gly 17

Molecular Characterization

V176
CD16a(Gly 17 - Gln 208)P08637-1
Avi
His

This protein carries an Avi tag (Avitag™) at the C-terminus, followed by a polyhistidine tag.

The protein has a calculated MW of 25.5 kDa. The protein migrates as 40-55 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

Biotinylation

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

Biotin:Protein Ratio

The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

 

Purity

>90% as determined by SDS-PAGE.

Formulation

Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.

Contact us for customized product form or formulation.

Reconstitution

Please see Certificate of Analysis for specific instructions.

For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

Storage

For long term storage, the product should be stored at lyophilized state at -20°C or lower.

Please avoid repeated freeze-thaw cycles.

No activity loss is observed after storage at:

  1. 4-8°C for 12 months in lyophilized state;
  2. -70°C for 3 months under sterile conditions after reconstitution.
 

SDS-PAGE

Biotinylated Human FCGR3A / CD16a (V176) (Catalog # CDA-H82E9) SDS-PAGE gel

Biotinylated Human FCGR3A / CD16a (V176) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity

Biotinylated Human FCGR3A / CD16a (V176) (Catalog # CDA-H82E9) ELISA bioactivity

(1) Immobilized Yervoy Ipilimumab at 5 μg/mL (100 μl/well) can bind Biotinylated Human FCGR3A / CD16a (V176) (Cat# CDA-H82E9) with a linear range of 0.03-0.25 μg/mL.

Request for protocol

(2) The bioactivity is measured by its binding ability to Ipilimumab in a SPR assay. Immobilized Biotinylated Human FCGR3A / CD16a (V176) Protein (Cat# CDA-H82E9) can bind Ipilimumab with affinity constant of around 158 nM (routinely tested).

 

Background

CD16 is a low affinity Fc receptor, and has been identified as Fc receptors FcγRIIIa (CD16a) and FcγRIIIb (CD16b). These receptors bind to the Fc portion of IgG antibodies. CD16 encoded by two different highly homologous genes in a cell type-specific manner.CD16 is found on the surface of natural killer cells, neutrophil polymorphonuclear leukocytes, monocytes and macrophages. CD16a antigen is also known as Low affinity immunoglobulin gamma Fc region receptor III-A, Fc-gamma RIII-alpha. CD16b is a low-affinity, GPI-linked receptor expressed by neutrophils and eosinophils, whereas CD16a is an intermediate affinity polypeptide-anchored transmembrane glycoprotein expressed natural killer cells, macrophages, subpopulation of T-cells, immature thymocytes and placentaltrophoblasts.CD16a is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody¬dependent cytotoxicity and clearance of immune complexes. Aberrant expression or mutations of CD16a is implicated in susceptibility to recurrent viral infections, systemic lupus erythematosus, and alloimmune neonatal neutropenia.

References

Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.


Limited Use License

The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.

Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.