CD16 Monoclonal Antibody (3G8), Super Bright™ 702, eBioscience™, Invitrogen™

Description

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 eBioCB16 (CB16) antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioCB16 (CB16) antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (0.06 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.

Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Super Bright 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 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 No. 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. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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: 702 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. CD16 (FCGR3A) is a 50-65 kDa cell surface molecule that exists in two forms - a transmembranous form expressed by NK cells and some T cells, and a phosphatidylinositol linked form expressed by granulocytes. CD16 is a low affinity receptor for IgG (FcR III), and is an important receptor mediating ADCC by NK cells. Human CD16 is expressed in two forms – FCGR3A and FCGR3B. FCGR3A is associated with the FcepsilonRI-gamma subunit and is responsible for antibody-dependent NK cell cytotoxicity. Mast cell FCGR3A is associated, with FcepsilonRI-beta subunit. Besides IgG, FCGR3A can be triggered also by oligomeric IgE. FCGR3B is a GPI-linked monomeric receptor expressed on neutrophils and is involved in their activation and induction of a pro-adhesive phenotype. Diseases associated with CD16 dysfunction include immunodeficiency 20 and systemic lupus erythematosus.