Anti-CD16b Rabbit Monoclonal Antibody for WB, FC, ELISA - O75015

Core Product Specifications and Parameters

Biological Background: CD16b Function and Localization

• CD16b (FcγRIIIB) is a low affinity receptor for the Fc region of immunoglobulins gamma, encoded by the FCGR3B gene. It is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein without a transmembrane domain, distinguishing it from FcγRIIIA.
• The receptor binds complexed or aggregated IgG and monomeric IgG, but unlike FcγRIIIA, it does not mediate antibody-dependent cellular cytotoxicity (ADCC) or phagocytosis, likely serving as a decoy receptor that traps immune complexes in the peripheral circulation without activating neutrophils.
• CD16b is expressed specifically by polymorphonuclear leukocytes (neutrophils) and by stimulated eosinophils, underscoring its role in innate immune surveillance on granulocytes.
• Subcellularly, CD16b is found on the cell membrane and can be released as a secreted form, mediated by cleavage of its GPI anchor.
• The protein contains two immunoglobulin-like domains and is extensively glycosylated; a predicted molecular weight of approximately 26 kDa (core protein 26,243 Da) is often observed as a diffuse band due to glycosylation.
• Key structural features include an N-terminal signal peptide (aa 1-18), the extracellular IgG-binding domain (aa 20-208, used as immunogen), and a C-terminal GPI-anchor attachment site. Multiple aliases (CD16, FCG3, FCGR3, FCR-10, FCRIII) reflect historical nomenclature.

Experimental Guidance and Technical Tips

• The immunogen covers the entire extracellular IgG-binding domain (aa 20-208) of human CD16b, which is well-suited for detecting both native and denatured protein across applications.
• For Western blot (WB), a band around 26 kDa is expected under reducing conditions; smearing or a diffuse band may occur due to glycosylation—validate with appropriate positive controls (e.g., neutrophil lysates) and consider deglycosylation if a sharp band is required.
• In flow cytometry (FC), use a compatible fluorophore-conjugated secondary antibody for detection of surface CD16b on live, unfixed neutrophils; avoid fixation methods that may alter GPI-anchored protein integrity.
• For ELISA, the antibody can be used as a capture or detection reagent; pairing with a non-competing anti-CD16b clone is recommended for sandwich formats, and assay conditions should be optimized with recombinant or purified CD16b protein.
• Cross-reactivity limited to human; no reactivity with mouse or rat CD16b is expected, so choose an appropriate model system or validate with human samples.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers and research labs with a comprehensive range of IVD raw materials, from validated antibody pairs and optimized monoclonal antibodies to ancillary reagents and rare-target sourcing. For CD16b and Fc receptor biology applications, our procurement services cover bulk quantities of this rabbit monoclonal antibody, expert curation of alternative antibody formats, and technical guidance to streamline your assay development.