Cytochrome P450 2C9 Inhibitor Screening Kit


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Cytochrome P450 2C9 Inhibitor Screening Kit

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Cat.No. : Kit-2117
Product Overview : CYP2C9 Inhibitor Screening Kit enables rapid screening of drugs and other new chemical entities (NCEs) for compound-CYP2C9 interaction in a reliable, high-throughput fluorescence-based assay. The kit provides a yeast microsomal preparation of human CYP2C9 and human cytochrome P450 reductase (CPR) enzymes. The assay utilizes a non-fluorescent CYP2C9 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 415/502 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96-well plate format.
Size : 100 assays
Applications : Rapid, high-throughput screening of drugs and novel ligands.Development of structure-activity relationship (SAR) models to predict CYP2C9 inhibition liability of novel compounds.Prediction of adverse drug-drug interaction potential and bioavailability for compounds metabolized by CYP2C9.
Storage : Store kit at -20°C and protected from light. Briefly centrifuge all small vials prior to opening. Allow the CYP2C9 Assay Buffer to warm to room temperature (RT) prior to use. Read entire protocol before performing the assay procedure.7-HFC Standard: Reconstitute in 110 µl of DMSO and vortex until fully dissolved to yield a 5 mM stock solution. The 7-HFC stock solution should be stored at -20°C and is stable for at least 3 freeze/thaw cycles.CYP2C9 Inhibitor (Sulfaphenazole): Reconstitute in 55 µl of acetonitrile and vortex until fully dissolved to yield a 20 mM stock solution. The stock solution is stable for 2 months at -20°C. To obtain a 300 µM working solution of sulfaphenazole (5X final concentration), add 15 µl of the 20 mM stock solution to 985 µl of CYP2C9 Assay Buffer. The 300 µM working solution should be stored at -20°C and used within one month.NADPH Generating System (100X): Reconstitute with 110 µl CYP2C9 Assay Buffer, aliquot and store at -20°C. Avoid repeated freeze/thaw cycles. Keep on ice while in use.β-NADP+ Stock (100X): Dissolve in 110 µl CYP2C9 Assay Buffer and vortex thoroughly (100X stock). Store at -20°C, stable for at least 3 freeze/thaw cycles.CYP2C9 Substrate: Reconstitute with 55 µl anhydrous HPLC-grade acetonitrile and vortex until fully dissolved to obtain a 10 mM stocksolution. Store at -20°C. When using the CYP2C9 Substrate stock solution, allow the vial to warm to RT before opening and promptlyretighten cap after use to avoid absorption of airborne moisture.Recombinant Human CYP2C9: The Recombinant Human CYP2C9 should be reconstituted immediately before use as directed in Protocol 2 below. Each vial is sufficient for preparation of 50 reactions in a 96-well plate format.

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