The antibody was purified by affinity chromatography, and conjugated with PerCP/Cy5.5 under optimal conditions. The solution is free of unconjugated PerCP/Cy5.5 and unconjugated antibody.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
FC
Recommended Usage:
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is 5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
* PerCP/Cy5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.
Application Notes:
Additional reported applications (for the relevant formats) include: immunohistochemical staining6 of acetone-fixed frozen tissue sections, immunoprecipitation3, stimulation4 of NK cell proliferation, blocking of phagocytosis5, and blocking of immunoglobulin binding to FcγRIII7,8. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 302014).
Application References:
1. Knapp, W., et al., Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York. 2. Schlossman, S., et al., Eds. 1995. Leucocyte Typing V. Oxford University Press. New York. 3. Edberg, J., et al., 1997. J. Immunol. 159:3849. 4. Hoshino, S., et al., 1991. Blood 78:3232. 5. Tamm, A., et al., 1996. Immunol. 157:1576. 6. Da Silva, D.M. et al., 2001. Int. Immunol. 13:633. 7. Holl, V., et al., 2004. J. Immunol. 173:6274. 8. Hober, D., et al., 2002. J. Gen. Virol. 83:2169. 9. Smed-Sörensen, A., et al., 2008. Blood 111:5037. (Block) PubMed 10. Timmerman, K.L., et al., 2008. J. Leukoc. Biol. 84:1271. (FC) PubMed
Human peripheral blood lymphocytes stained with 3G8 PerCP/Cy5.5
Description:
CD16 is known as low affinity IgG receptor III (FcγRIII). It is expressed as two distinct (CD16a and CD16b) forms. CD16a (FcγRIIIA) is a 50-65 kD polypeptide-anchored transmembrane protein. It is expressed on the surface of NK cells, activated monocytes, macrophages, and placental trophoblasts in humans. CD16b (FcγRIIIB) is a 48 kD glycosylphosphatidylinositol (GPI)-anchored protein. It is over 95% homologous to CD16a in extracellular domain and is expressed specifically on neutrophils. CD16 binds aggregated IgG or IgG-antigen complex that functions in NK cell activation, phagocytosis, and antibody-dependent cell-mediated cytotoxicity (ADCC). The 3G8 antibody blocks neutrophil phagocytosis and stimulates NK cell proliferation.
Other Names:
FcγRIII
Structure:
Ig superfamily, transmembrane form (50-65 kD) or GPI-linked form, 48 kD
Distribution:
NK cells, activated monocytes, macrophages, neutrophils
Function:
Low affinity IgG Fc receptor, phagocytosis, ADCC
Ligand Receptor:
Aggregated IgG, IgG-antigen complex
Antigen References:
1. Fleit, H., et al., 1982. PNAS 79: 3275. 2. Stroncek, D., et al., 1991. Blood 77: 1572. 3. Wirthmueller, U., et al., 1992. J. Exp. Med. 175: 1381.
