Biolegend, 302049, Ultra-LEAF™ Purified anti-human CD16 Antibody, 100μg

CD16 is known as low affinity IgG receptor III (FcγRIII). It is expressed as two distinct forms (CD16a and CD16b). 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. Its extracellular domain is over 95% homologous to that of CD16a, and it is expressed specifically on neutrophils. CD16 binds aggregated IgG or IgG-antigen complex which functions in NK cell activation, phagocytosis, and antibody-dependent cell-mediated cytotoxicity (ADCC).
100μg
Verified Reactivity: Human, Cynomolgus, Rhesus
Reported Reactivity: African Green, Baboon, Capuchin Monkey, Chimpanzee, Common Marmoset, Pigtailed Macaque, Sooty Mangabey, Squirrel Monkey
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human PMN cells
Formulation: 0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.
Endotoxin Level: Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.
Preparation: The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration: The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.
Application: FC - Quality testedCyTOF® - VerifiedBlock, IHC-F, IP, PG, Stim - Reported in the literature, not verified in house
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤2.0 µg per million cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: The 3G8 antibody clone blocks neutrophil phagocytosis and stimulates NK cell proliferation. It has been reported that this clone interacts with the FcγRIIa and FcγRIIIb receptors causing neutrophil activation and aggregation18. Due to this phenomenon staining in whole blood may cause a reduction in the number of granulocytes or alter their scatter profile.Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections6, immunoprecipitation3, stimulation of NK cell proliferation4, blocking of phagocytosis5, and blocking of immunoglobulin binding to FcγRIII7,8. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 302049, 302050, 302057, 302058).
Application References(PubMed link indicates BioLegend citation): Knapp W, et al. Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York. Schlossman S, et al. Eds. 1995. Leucocyte Typing V. Oxford University Press. New York. Edberg J, et al. 1997. J. Immunol. 159:3849. (IP) Hoshino S, et al. 1991. Blood 78:3232. (Stim) Tamm A, et al. 1996. Immunol. 157:1576. (Block) Da Silva DM, et al. 2001. Int. Immunol. 13:633. (IHC) Holl V, et al. 2004. J. Immunol. 173:6274. (Block) Hober D, et al. 2002. J. Gen. Virol. 83:2169. (Block) Brainard DM, et al. 2009. J. Virol. 83:7305. PubMed Smed-Sörensen A, et al. 2008. Blood 111:5037. (Block) PubMed Timmerman KL, et al. 2008. J. Leukoc. Biol. 84:1271. (FC) PubMed Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Rout N, et al. 2010. PLoS One 5:e9787. (FC) Kim WK, et al. 2006. Am. J. Pathol. 168:822. (FC) Boltz A, et al. 2011. J. Biol Chem. 286:21896. PubMed Wu Z, et al. 2013. J. Virol. 87:7717. PubMed Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG) Vossebeld PJ, et al. 1997. Biochem J. 323:87-94 (Stim)
Product Citations: Yamin R, et al. 2021. Nature. 599:465. PubMed Vidard L, et al. 2023. Immun Inflamm Dis. 11:e749. PubMed Cluff E, et al. 2022. Cancer Immunol Immunother. . PubMed Waight JD, et al. 2018. Cancer Cell. 33:1033. PubMed Zhu C, et al. 2020. Front Immunol. 1.688194444. PubMed Gimpel AK, et al. 2022. Front Immunol. 12:761816. PubMed Beasley AB, et al. 2021. Methods Mol Biol. 2265:213. PubMed Thulin NK, et al. 2020. Cell Reports. 31(6):107642. PubMed
RRID: AB_2783156 (BioLegend Cat. No. 302049) AB_2561481 (BioLegend Cat. No. 302050) AB_2572005 (BioLegend Cat. No. 302057) AB_2572006 (BioLegend Cat. No. 302058)
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
Cell Type: Dendritic cells, Macrophages, Monocytes, Neutrophils, NK cells
Biology Area: Immunology, Innate Immunity
Molecular Family: CD Molecules, Fc Receptors
Antigen References: 1. Fleit H, et al. 1982. P. Natl. Acad. Sci. USA 79:3275. 2. Stroncek D, et al. 1991. Blood 77:1572. 3. Wirthmueller U, et al. 1992. J. Exp. Med. 175:1381.
Gene ID: 2214
UniProt: View information about CD16 on UniProt.org
Clone: 3G8
Regulatory Status: RUO
Workshop: V NK80
Other Names: FcγRIII, Fc gamma receptor, Fc gamma receptor 3
Isotype: Mouse IgG1, κ
Q: Is our human Trustain FcX™ (cat# 422302) compatible with anti human CD16, CD32 and CD64 clones 3G8, FUN-2 and 10.1 respectively?
A: Yes
Q: Do you guarantee that your antibodies are totally pathogen free?
A: BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.
Q: Does BioLegend test each Ultra-LEAF™ antibody by functional assay?
A: No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.
Q: Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?
A: No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.
Q: Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?
A: We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.