CD16 Monoclonal Antibody (eBioCB16 (CB16)), APC-eFluor™ 780, eBioscience™

Product Specific Information

Description: The eBioCB16 monoclonal antibody recognizes CD16 (Fc gammaRIII), the low-affinity receptor for IgG with an apparent molecular weight of 50-80 kDa. CD16 is represented by two similar genes, CD16A (Fc gammaRIIIA), which exists as a hetero-oligomeric polypeptide-anchored form in macrophages and NK cells and CD16B (Fc gammaRIIIB), which exist as a monomeric GPI-anchored form in neutrophils. Furthermore, there are two known polymorphisms of CD16B, NA-1 and NA-2. Individuals homozygous for NA-2 show a lower phagocytic capacity compared with NA-1. CD16 binds IgG in the form of immune complexes and shows preferential binding of IgG1 and IgG3 isotypes and minimal binding of IgG2 and IgG4. Upon IgG binding, both CD16 isoforms initiate signal transduction cascades that lead to a variety of responses including antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, degranulation and proliferation.

Applications Reported: This CB16 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This eBioCB16 (CB16) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

APC-eFluor 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.

Light sensitivity: This tandem is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

CD16, known as Fc gamma receptor III, exists in two distinct isoforms: Fc gamma RIIIA and Fc gamma RIIIB. These isoforms are encoded by the genes FCGR3A and FCGR3B, respectively, which are located on chromosome 1 and share a high degree of similarity. Fc gamma RIIIA is expressed as a polypeptide-anchored form on monocytes, macrophages, and lymphocytes such as natural killer (NK) cells, but not on T or B cells. This receptor recognizes the Fc portion of immunoglobulin G (IgG) and is involved in the clearance of immune complexes from circulation. Upon binding IgG, Fc gamma RIIIA activates signal transduction pathways that result in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, cytokine release, and antigen presentation. Fc gamma RIIIB is expressed on polymorphonuclear neutrophils (PMN) as a glycosylphosphatidylinositol (GPI)-anchored form. It plays a role in similar immune functions, including cellular-mediated cytotoxicity and enhancement of virus infections. Mutations in the FCGR3A gene have been associated with Immunodeficiency 20 and linked to susceptibility to recurrent viral infections, systemic lupus erythematosus, and alloimmune neonatal neutropenia. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. Diseases associated with FCGR3A include Immunodeficiency 20 and Herpes Zoster.