Promega

ADCC Reporter Bioassay, Core Kit

Varenummer: G7010
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Fc receptor-mediated antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action (MOA) by which antibodies target disease cells for elimination. Classic methods used to measure ADCC utilize primary donor peripheral blood mononuclear cells (PBMCs) or purified natural killer (NK) cells that express Fc receptors on their cell surface. Following engagement with the Fc region of a relevant antibody bound to a target disease cell, Fc receptors transduce intracellular signals within the effector cell resulting in elimination of the target cell. These primary cell-based assays are highly variable as a result of donor differences and the requirement for cell culture and expansion. The ADCC Reporter Bioassay provides a biologically relevant and specific MOA-based measure of ADCC without the complex workflow and variability inherent in primary cell-based assays. Specifically, primary donor PBMC or NK cells are replaced with a Jurkat cell stably expressing human FcgammaRIIIa (either the high-affinity V158 or low-affinity F158 receptor) and NFAT-induced luciferase. Importantly, the ADCC Reporter Bioassay demonstrates rank ordering of antibody activity equivalent to classic LDH release ADCC bioassays. The bioassay also can be used to quantify effects of antibody glycosylation on Fc effector function. The ADCC Reporter Bioassay is available in multiple product kit formats: Complete Kits (Include ADCC Bioassay Effector Cells, Target Cells, Control Antibody, Cell Culture Medium and Assay Reagents. Recommended for use as a starter kit. Available with either Raji or WIL2-S Target Cells.) Core Kits (Include ADCC Bioassay Effector Cells, Cell Culture Medium and Assay Reagents. Recommended for routine use with customer-defined antibody and target cells. Available in 1X and 5X sizes.) Target Kits (Include ADCC Bioassay Target Cells and Control Antibody. Recommended for use as a control with all Fc Effector Bioassay Core Kits. Available with either Raji or WIL2-S Target Cells.)|

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Note: The ADCC Reporter Bioassay components are shipped separately because of temperature requirements. The ADCC Bioassay Effector Cells and Target Cells are shipped on dry ice. The Bio-Glo(TM); Luciferase Assay System and Low IgG Serum are shipped on dry ice, separately from the cells. The RPMI 1640 Medium is shipped at ambient temperature. The Control Ab, Anti-CD20, is shipped on gel ice.
Upon arrival, immediately transfer the vials of ADCC Bioassay Effector Cells and Target Cells for long-term storage below -140 C (freezer or liquid nitrogen vapor phase). The cells are sensitive, and care should be taken when handling. For safety reasons do not store cell vials submerged in liquid nitrogen. Store the Low IgG Serum at -20 C. Avoid multiple freeze-thaw cycles. Store the Control Ab, Anti-CD20, at 4 C. Store the Bio-Glo(TM); Luciferase Assay Buffer and Luciferase Assay Substrate at -20 C. For optimal performance, use reconstituted Bio-Glo(TM); Luciferase Assay Reagent on the day of preparation. However, once reconstituted, you can store Bio-Glo(TM); Luciferase Assay Reagent at -20 C for up to 6 weeks. Store RPMI 1640 Medium at 4 C protected from fluorescent light.|

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Christine Rindal Ibra 944 34 009 christine.rindal.ibra@nmas.no
Claudia Emmanuel 951 51 950 claudia.emmanuel@nmas.no
Monica Laukas 404 40 960 monica.laukas@nmas.no

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