Staining of normal human (left) and rhesus (right) peripheral blood cells with Mouse IgG1 κ Isotype Control PerCP-eFluorŪ 710 (cat. 46-4714) (blue histogram) or Anti-Human/Non-Human Primate CD16 PerCP-eFluorŪ 710. Cells in the lymphocyte (purple histogram) and granulocyte (orange histogram) gates were used for analysis.
Contents: Anti-Human/Non-Human Primate CD16 PerCP-eFluorŪ 710 Catalog Number: 46-0166 Concentration: 5 µl (0.125 µg)/test Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. Do not freeze. Light-sensitive material. Clone: 3G8 Host/Isotype: Mouse IgG1
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46-0166-41
25 tests
46-0166-42
100 tests
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa FluorŪ 647 conjugated products with eFluorŪ 660 conjugated products.
Description
This 3G8 monoclonal antibody reacts with human and non-human primate CD16, which is also known as the low-affinity FcγRIII. CD16 exists as two distinct isoforms, FcγRIIIA and FcγRIIIB. In humans, FcγRIIIA 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. FcγRIIIB 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
For research use only, not for diagnostic or therapeutic use. This 3G8 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This 3G8 antibody has been pre-titrated and tested by flow cytometric analysis of normal human and rhesus peripheral blood cells. This can 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 105 to 108 cells/test.
PerCP-eFluor® 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative.
Our testing indicates that PerCP-eFluor® 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols.
Choi EI, Wang R, Peterson L, Letvin NL, Reimann KA. Use of an anti-CD16 antibody for in vivo depletion of natural killer cells in rhesus macaques. Immunology. 2008 Jun;124(2):215-22. (3G8, FA, FC)
Rogers KA, Scinicariello F, Attanasio R. IgG Fc receptor III homologues in nonhuman primate species: genetic characterization and ligand interactions. J Immunol. 2006 Sep 15;177(6):3848-56. (3G8, FC)
Naziruddin B, Duffy BF, Tucker J, Mohanakumar T. Evidence for cross-regulation of Fc gamma RIIIB (CD16) receptor-mediated signaling by Fc gamma RII (CD32) expressed on polymorphonuclear neutrophils. J Immunol. 1992 Dec 1;149(11):3702-9. (3G8, FA)
Perussia B, Trinchieri G. Antibody 3G8, specific for the human neutrophil Fc receptor, reacts with natural killer cells. J Immunol. 1984 Mar;132(3):1410-5. (3G8, FC)
