Fc gamma RIIIA/CD16a Antibody (DJ130c) [CoraFluor™ 1]

CoraFluor™ 1, amine reactive (Catalog:7920) and CoraFluor™ 2, amine reactive (Catalog # 7950) are terbium-based probes that have been developed for use as TR-FRET donors. They emit wavelengths compatible with commonly used fluorescent acceptor dyes such as BODIPY® (or BDY) and Janelia Fluor® dyes, FITC (Catalog # 5440), TMR and Cyanine 5 (Catalog # 5436). CoraFluor™ fluorescence is brighter and more stable in biological media than existing TR-FRET donors, leading to enhanced sensitivity and improved data generation. CoraFluor™ 1 exhibits excitation upon exposure to a 337 nm UV laser.

• Reactivity: Hu
• Applications: Flow, IHC
• Clone: DJ130c
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: CoraFluor 1

Fc gamma RIIIA/CD16a Antibody (DJ130c) [CoraFluor™ 1] Summary

• Description: CoraFluor(TM) 1 is a high performance terbium-based TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer) or TRF (Time-Resolved Fluorescence) donor for high throughput assay development. CoraFluor(TM) 1 absorbs UV light at approximately 340 nm, and emits at approximately 490 nm, 545 nm, 585 nm and 620 nm. It is compatible with common acceptor dyes that absorb at the emission wavelengths of CoraFluor(TM) 1. CoraFluor(TM) 1 can be used for the development of robust and scalable TR-FRET binding assays such as target engagement, ternary complex, protein-protein interaction and protein quantification assays.
  
  CoraFluor(TM) 1, amine reactive
  
  CoraFluor(TM) 1, thiol reactive
  
  For more information, please see our CoraFluor(TM) TR-FRET technology flyer.
• Immunogen: CD34 antigen
• Specificity: NB100-64346 recognizes human CD16, a 50-65kD cell surface molecule, which is the low affinity receptor for IgG (FcR III). CD16 exists as a transmembranous form (Fc gammaRIIIA, or CD16A) and a glycosyl phosphatidylinositol (GPI) anchored form (Fc gammaRIIIB, or CD16B). CD16A is expressed by NK cells, some T cells, and macrophages, whereas CD16B is primarily expressed by granulocytes. In addition, CD16B exists as two allelic variants NA1 and NA2. DJ130c recognizes all polymorphonuclear cells irrespective on their NA phenotype. Studies show that the DJ130c antibody recognizes an epitope in the first membrane-distal domain of CD16 and recognizes both CD16A and CD16B.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: FCGR3A
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Reactivity Notes

Predicted cross-reactivities: Macaque

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark. Do not freeze.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein A or G purified

Notes

CoraFluor (TM) is a trademark of Bio-Techne Corp. Sold for research purposes only under agreement from Massachusetts General Hospital. US patent 2022/0025254

Alternate Names for Fc gamma RIIIA/CD16a Antibody (DJ130c) [CoraFluor™ 1]

• 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 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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

FAQs for Fc gamma RIIIA/CD16a Antibody (NB100-64346CL1). (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

• Gene Symbol: FCGR3A