CD16 Monoclonal Antibody (3G8), Super Bright™ 600, eBioscience™

Product Specific Information

Description: This 3G8 monoclonal antibody reacts with human and non-human primate CD16, which is also known as the low-affinity Fc gamma RIII. CD16 exists as two distinct isoforms, Fc gamma RIIIA and Fc gamma RIIIB. In humans, Fc gamma RIIIA is expressed as a polypeptide-anchored form on monocytes, macrophages, and lymphocytes such as NK cells. T and B cells do not express this Fc receptor. Fc gamma RIIIB is also detected on neutrophils as a GPI-anchored form. Expression of CD16 on lymphocytes and monocytes is similar in non-human primates. However, while CD16 is not found on neutrophils in macaques and baboons, this receptor is detected on these cells in sooty mangabeys. Binding of IgG leads to activation of signal transduction pathways, resulting in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, cytokine release, and antigen presentation.

This monoclonal antibody has been reported to have several functional activities, including inhibition of cytotoxic ability, activation of cell signaling, and NK cell depletion in vivo. Moreover, the 3G8 antibody clone has been demonstrated to work on capuchin monkey, chimpanzee, common marmoset, cynomologous monkey, hamadyras baboon, olive baboon, pigtailed macaque, rhesus, and squirrel monkey.

Based on cross-blocking studies 3G8 recognizes the same epitope as CB16. However, 3G8 and B73.1 antibody clones bind distinct epitopes.

Applications Reported: This 3G8 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 3G8 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.125 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.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

Light sensitivity: This tandem dye 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 of cell sample + 100 µL of 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: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

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.