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Fc gamma RIII (CD16) Antibody (3G8) - Chimeric

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-21452

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Human, Baboon, Chimpanzee, Cynomolgus Monkey, Rhesus Macaque

Applications

Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Surface Plasmon Resonance (SPR)

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # 3G8

Concentration

1 mg/ml

Product Specifications

Immunogen

Human PMN (polymorphonuclear leukocytes) cells.

Reactivity Notes

Expected to react with Pigtailed Macaque, Capuchin Monkey, Squirrel Monkey, Sooty Mangabey, Cotton-topped Tamarin, Marmoset.

Specificity

This antibody binds to CD16 at an epitope which is not the IgG-binding site. This antibody cannot distinguish between CD16a transmembrane isoform found on the surface of NK cells and macrophages (which can trigger the cytolytic response), and the GPI-linked CD16b isoform present on neutrophils or soluble CD16 found in the plasma. CD16 is the low affinity IgG receptor III and binds to aggregated IgG or IgG-antigen complex which functions in NK cell activation, phagocytosis and ADCC (antibody-dependent cell-mediated cytotoxicity).

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Description

This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.

Applications for Fc gamma RIII (CD16) Antibody (3G8) - Chimeric

Application
Recommended Usage

Block/Neutralize

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Surface Plasmon Resonance (SPR)

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This antibody was used to construct a recombinant bispecific scFv (bsscFv) along with the mAb clone F3.3 anti-HLA class II antibody (connected by a (Gly4Ser)4 linker)which was expressed in SF21 insect cells. This bsscFv allows NK cells to target malignant B lymphoid cells and mediates effector cell lysis (ASCC) of the malignant cells (also lysis of primary leukaemia samples). 3G8 is also able to block neutrophil phagocytosis and stimulates NK cell proliferation, and can be used in FC, SPR, IP and IHC studies.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Fc gamma RIII (CD16)

Fc gamma receptor III (RIII), also called CD16, 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 RIII (CD16) is one of three subclasses of Fc gamma receptors which includes Fc gamma RI (CD64) and Fc gamma RII (CD32) (1,2). The two forms 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, while human Fc gamma RIIIB protein is 233 aa long with a calculated MW of 26 kDa (1,4,5). Each protein contains between five to six glycosylation sites (1,4,5).

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). Fc gamma RIIIB/CD16b is primarily expressed on neutrophils as a glycosylphosphatidylinositol (GPI)-anchored protein but is also expressed on a subset of basophils and can be induced on eosinophils (1-3). The soluble form of CD16 (sCD16) which 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 (6). 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 (6). CD16a is also capable of antibody dependent cellular cytotoxicity (ADCC) through recognition of antibodies bound to target cells (6-7). 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 (7). 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 (6-7).

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. Uniprot (O75015)

6. 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

7. 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

Alternate Names

FcgRIII

Gene Symbol

FCGR3A

UniProt

Additional Fc gamma RIII (CD16) Products

Product Documents for Fc gamma RIII (CD16) Antibody (3G8) - Chimeric

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Fc gamma RIII (CD16) Antibody (3G8) - 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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