{"product_id":"biolegend-285021","title":"Biolegend, 285021, Spark Blue™ 574 anti-human CD16 (Flexi-Fluor™) Antibody, 50μg","description":"\u003cp\u003eCD16 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).\u003cbr\u003e\n50μg\u003cbr\u003e\nVerified Reactivity: Human, Cynomolgus, Rhesus\u003cbr\u003e\nReported Reactivity: African Green, Baboon, Capuchin Monkey, Chimpanzee, Common Marmoset, Pigtailed Macaque, Sooty Mangabey, Squirrel Monkey\u003cbr\u003e\nAntibody Type: Monoclonal\u003cbr\u003e\nHost Species: Mouse\u003cbr\u003e\nImmunogen: Human PMN cells\u003cbr\u003e\nFormulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide\u003cbr\u003e\nPreparation: The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.\u003cbr\u003e\nConcentration: 0.2 mg\/mL\u003cbr\u003e\nStorage \u0026amp; Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.\u003cbr\u003e\nApplication: FC\u003cbr\u003e\nRecommended Usage: Flexi-Fluors™ are provided at a standard 0.2 mg\/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg\/mL.For example, make five 1:1 serial dilutions of the 0.2 mg\/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells\/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. * Spark Blue™ 574 has a maximum excitation of 506 nm and a maximum emission of 574 nm.\u003cbr\u003e\nExcitation Laser: Blue Laser (488 nm)\u003cbr\u003e\nApplication 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 \u0026lt; 0.01 EU\/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 302049, 302050, 302057, 302058).\u003cbr\u003e\nAdditional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.\u003cbr\u003e\nApplication 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)\u003cbr\u003e\nRRID: AB_3106286 (BioLegend Cat. No. 285021)\u003cbr\u003e\nStructure: Ig superfamily, transmembrane form (50-65 kD) or GPI-linked form (48 kD)\u003cbr\u003e\nDistribution: NK cells, activated monocytes, macrophages, neutrophils\u003cbr\u003e\nFunction: Low affinity IgG Fc receptor, phagocytosis, ADCC\u003cbr\u003e\nLigand\/Receptor: Aggregated IgG, IgG-antigen complex\u003cbr\u003e\nCell Type: Dendritic cells, Macrophages, Monocytes, Neutrophils, NK cells\u003cbr\u003e\nBiology Area: Immunology, Innate Immunity\u003cbr\u003e\nMolecular Family: CD Molecules, Fc Receptors\u003cbr\u003e\nAntigen 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.\u003cbr\u003e\nGene ID: 2214\u003cbr\u003e\nUniProt: View information about CD16 on UniProt.org\u003cbr\u003e\nClone: 3G8\u003cbr\u003e\nRegulatory Status: RUO\u003cbr\u003e\nWorkshop: V NK80\u003cbr\u003e\nOther Names: FcγRIII, Fc gamma receptor, Fc gamma receptor 3\u003cbr\u003e\nIsotype: Mouse IgG1, κ\u003cbr\u003e\nQ: Is our human Trustain FcX™ (cat# 422302) compatible with anti human CD16, CD32 and CD64 clones 3G8, FUN-2 and 10.1 respectively?\u003cbr\u003e\nA: Yes\u003cbr\u003e\nQ: What are Flexi-Fluors?\u003cbr\u003e\nA: Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.\u003cbr\u003e\nQ: How quickly will I receive my order?\u003cbr\u003e\nA: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.\u003cbr\u003e\nQ: How are Flexi-Fluors different from regular catalog products?\u003cbr\u003e\nA: Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.\u003cbr\u003e\nQ: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?\u003cbr\u003e\nA: Flexi-Fluors are provided at a standard 0.2 mg\/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg\/mL. \u003c\/p\u003e\n\n\u003cp\u003e \u003c\/p\u003e\n\n\u003cp\u003eFor example, make five 1:1 serial dilutions of your 0.2 mg\/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells\/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.\u003cbr\u003e\nQ: I can’t find the antibody-dye combination that I need. When will it be available?\u003cbr\u003e\nA: We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.\u003cbr\u003e\nQ: I need help to validate the performance of my Flexi-Fluor. Who should I contact?\u003cbr\u003e\nA: Please get in touch with Technical Service for assistance.\u003cbr\u003e\nQ: Can I order more than 50 μg of a Flexi-Fluor?\u003cbr\u003e\nA: Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.\u003cbr\u003e\nQ: What is the expiration date of my Flexi-Fluor?\u003cbr\u003e\nA: Expiration dates can be found on the vial label or by using our CoA lookup tool.\u003cbr\u003e\nQ: What if my requested antibody-dye combination doesn't perform as expected?\u003cbr\u003e\nA: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.\u003c\/p\u003e","brand":"Biolegend","offers":[{"title":"Default Title","offer_id":46862398521513,"sku":"285021","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/ar\/products\/biolegend-285021","provider":"Iright","version":"1.0","type":"link"}