Description
Early screening of a drug candidate’s potential to induce or inhibit CYP2B6 is a critical step in the drug development process. Cayman’s CYP2B6 Induction Reporter Assay Kit is a novel method to evaluate human Constitutive Androstane Receptor 3 (hCAR3)-mediated CYP2B6 induction for the discovery of hepatic CYP inducers. Our Reverse Transfection Reporter Assays have overcome many of the disadvantages of other transfection approaches. Cayman’s CYP2B6 Induction Reporter Assay Kit consists of a 96-well plate coated with a transfection complex containing a Secreted Alkaline Phosphatase (SEAP) reporter regulated by the human CYP2B6 gene promoter. The complex also contains two nuclear receptor expression constructs, human Constitutive Androstane Receptor 3 (hCAR3) and human Hepatocyte Nuclear Factor 4α (HNF4α). Cells grown on the CYP2B6 Reverse Transfection Strip Plate will introduce the reporter gene and express CAR3 and HNF4α. Binding of inducer-activated receptor and transcription factors to the CYP2B6 promoter initiates a signal resulting in expression of SEAP, which is secreted into the cell culture medium. Aliquots of medium are removed at time intervals beginning at about 24 hours and SEAP activity is measured simply by adding a luminescence-based alkaline phosphatase substrate provided in the kit. The kit is simple to use and can be easily adapted to high-throughput screening for potential CAR-mediated CYP2B6 inducers. A known CYP2B6 inducer, CITCO, is included in the kit for use as a positive control.
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Application|Reverse Transfection Reporter Assay||Product Type|Assay Kits|Cell-Based Assays|Reverse Transfection||Research Area|Toxicology|Drug Metabolism|Cytochrome P450
