Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

Novus Biologicals | Catalog # NBP3-00383

Novus Biologicals
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Key Product Details

Sample Type & Volume Required Per Well

Serum, plasma, cell supernatant, and other biological fluids (100 ul)

Sensitivity

4.69 ng/mL

Assay Range

7.81 - 500 ng/mL
View all Fc gamma RIIIA/CD16a ELISA Kits »

Product Specifications

Assay Type

Sandwich-ELISA

Kit Type

ELISA Kit (Colorimetric)

Species

Human

Specificity

This kit recognizes Human Fc gamma RIIIA/CD16a in samples. No significant cross-reactivity or interference between Human Fc gamma RIIIA/CD16a and analogues was observed.

Precision

Intra-Assay Precision (Precision within an assay) 5.67%

Inter-Assay Precision (Precision between assays) 5.61%

Recovery for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

Recovery

88 - 108%

Linearity

Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

Scientific Data Images for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

ELISA: Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric) [NBP3-00383] -

ELISA: Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric) [NBP3-00383] - Standard Curve Reference

Kit Contents for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

  • Biotinylated Detection Ab Diluent
  • Concentrated Biotinylated Detection Ab (100x)
  • Concentrated HRP Conjugate (100x)
  • Concentrated Wash Buffer (25x)
  • HRP Conjugate Diluent
  • Micro ELISA Plate (Dismountable)
  • Plate Sealer
  • Product Manual
  • Reference Standard
  • Sample Diluent
  • Stop Solution
  • Substrate Reagent

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage of components varies. See protocol for specific instructions.

Background: Fc gamma RIIIA/CD16a

Fc gamma receptor IIIA (RIIIA), also called CD16a, is an activating natural killer (NK) cell receptor that binds the Fc portion of immunoglobulin G (IgG) antibodies and is responsible for eliciting a host defense against microbial pathogens (1). Fc gamma RIIIA/CD16a belongs to the Fc gamma RIII (CD16) subclass of Fc gamma receptors (1,2). The two other subclasses of receptors are Fc gamma RI (CD64) and Fc gamma RII (CD32) (1,2). The two form of Fc gamma RIII are Fc gamma RIIIA (CD16a) and Fc gamma RIIIB (CD16b), which are encoded by two different homologous genes, FCGR3A and FCGR3B, respectively (1-3). The human Fc gamma RIIIA protein is 254 amino acids (aa) in length with a theoretical molecular weight (MW) of 29 kDa (1,4,5). CD16a contains five glycosylation sites, two disulfide bonds, and two Ig-like C2 domains (4). Fc gamma RIIIA/CD16a is expressed as a transmembrane protein on NK cells and on a subset of monocytes, macrophages, CD4+ T cells, basophils, and mast cells (1-3). The soluble form of CD16 (sCD16) is often produced following exposure to inflammatory signals and protein shedding via metalloproteinases (3). Reduced sCD16 levels have been found in patients with multiple myeloma (3).

Activating NK cell receptor function has been harnessed for its potential in tumor immunotherapy (5). One immunotherapy strategy is using bi- and tri-specific NK cell engagers (BiKE and TriKE) to target the Fc gamma RIIIA/CD16a receptor with tumor-associated antigens to stimulate a cytotoxic response and mount an attack on tumor cells (5). CD16a is also capable of antibody dependent cellular cytotoxicity (ADCC) through recognition of antibodies bound to target cells (5-6). CD16-induced NK cell activation allows for NK co-receptor expression including stimulatory receptors like CD137 or inhibitory receptors like TIGIT and PD-1, which serve as additional regulatory checkpoints during ADCC (6). Therapeutic antibodies for cancer treatment like rituximab or trastuzumab can be recognized by Fc gamma RIIIA/CD16a to activate NK cell-mediated killing of tumor cells (5-6).

References

1. Fossati, G., Bucknall, R. C., & Edwards, S. W. (2001). Fcgamma receptors in autoimmune diseases. European Journal of Clinical Investigation, 31(9), 821-831. https://doi.org/10.1046/j.1365-2362.2001.00881.x

2. Patel, K. R., Roberts, J. T., & Barb, A. W. (2019). Multiple Variables at the Leukocyte Cell Surface Impact Fc gamma Receptor-Dependent Mechanisms. Frontiers in Immunology, 10, 223. https://doi.org/10.3389/fimmu.2019.00223

3. Moldovan, I., Galon, J., Maridonneau-Parini, I., Roman Roman, S., Mathiot, C., Fridman, W. H., & Sautes-Fridman, C. (1999). Regulation of production of soluble Fc gamma receptors type III in normal and pathological conditions. Immunology Letters, 68(1), 125-134. https://doi.org/10.1016/s0165-2478(99)00041-3

4. Uniprot (P08637)

5. Sivori, S., Pende, D., Quatrini, L., Pietra, G., Della Chiesa, M., Vacca, P., Tumino, N., Moretta, F., Mingari, M. C., Locatelli, F., & Moretta, L. (2021). NK cells and ILCs in tumor immunotherapy. Molecular Aspects of Medicine, 80, 100870. https://doi.org/10.1016/j.mam.2020.100870

6. Muntasell, A., Ochoa, M. C., Cordeiro, L., Berraondo, P., Lopez-Diaz de Cerio, A., Cabo, M., Lopez-Botet, M., & Melero, I. (2017). Targeting NK-cell checkpoints for cancer immunotherapy. Current Opinion in Immunology, 45, 73-81. https://doi.org/10.1016/j.coi.2017.01.003

Long Name

Fc gamma Receptor III A

Alternate Names

CD16a, FCGR3A, FcgRIIIA

Gene Symbol

FCGR3A

Additional Fc gamma RIIIA/CD16a Products

Product Documents for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

Certificate of Analysis

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Product Specific Notices for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

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FAQs for Human Fc gamma RIIIA/CD16a ELISA Kit (Colorimetric)

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  • Q:

    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.

    A:

    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.

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