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.

Name: Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric
Brand: Novus Biologicals
SKU: NBP3-48293

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.

Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-48293

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Recombinant Monoclonal Antibody

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species Reactivity

Human

Applications

ELISA, Flow Cytometry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Human IgG₁ Clone # DMC219

Concentration

LYOPH mg/ml

View all Fc gamma RIIIA/CD16a Antibodies »

Product Specifications

Immunogen

Fc gamma RIIIA/CD16a recombinant protein expressed in mammalian cells (Accession# P08637). The exact amino acid sequence is proprietary.

Clonality

Monoclonal

Host

Human

Isotype

IgG₁

Description

This product is shipped at ambient temperature. Store at -20C to -80C for 12 months in lyophilized form. After reconstitution, if not intended for use within a month, aliquot and store at -80C (Avoid repeated freezing and thawing).

Scientific Data Images for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Flow Cytometry: Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric [NBP3-48293]

Flow cytometry analysis with Anti-Fc gamma RIIIA/CD16a (DMC219) on HEK293 cells transfected with human Fc gamma RIIIA/CD16a (Blue histogram) or HEK293 transfected with irrelevant protein (Red histogram).

Applications for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Application

Recommended Usage

ELISA

1:5000-10000

Flow Cytometry

1:100

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified from cell culture supernatant

Reconstitution

Dissolve the lyophilized antibody in sterile and low endotoxin water. The recommend volume of water is lot specific. Please refer to the instructions printed on the product vial or contact technical support.

Formulation

PBS, pH 7.4, 5-8% Trehalose

Preservative

Trehalose

Concentration

LYOPH mg/ml

Shipping

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

Stability & Storage

Store at -20C in powder form. Store at -80C once reconstituted.

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 Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

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COA

Product Specific Notices for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

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.

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Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-48293

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

Concentration

View all Fc gamma RIIIA/CD16a Antibodies »

Product Specifications

Immunogen

Clonality

Host

Isotype

Description

Scientific Data Images for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Flow Cytometry: Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric [NBP3-48293]

Applications for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

ELISA

Flow Cytometry

Formulation, Preparation, and Storage

Purification

Reconstitution

Formulation

Preservative

Concentration

Shipping

Stability & Storage

Background: Fc gamma RIIIA/CD16a

Long Name

Alternate Names

Gene Symbol

Additional Fc gamma RIIIA/CD16a Products

Product Documents for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

Product Datasheets

COA

Certificate of Analysis

Product Specific Notices for Fc gamma RIIIA/CD16a Antibody (DMC219) - Chimeric

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