Highly Sensitive; Includes human CYP2C9 - NADPH Reductase as positive control.
- Simple, convenient, highly sensitive
- Fluorescent assay enables easy determination of CYP2C9 activity in a variety of biological samples
- The substrate shows low background and a high signal-to-noise ratio
- Kit includes CYP2C9 inhibitor (Tienilic Acid) and a stable, recombinant human CYP2C9 co-expressed with NADPH Reductase as a positive control
Cytochrome P450 2C9 (CYP2C9, EC 1.14.14.1) is a member of the cytochrome P450 monooxidase (CYP) family of microsomal xenobiotic metabolism enzymes. CYP2C9 is primarily expressed in the liver and catalyzes oxidation of small, weakly acidic or hydrophobic molecules containing an aromatic moiety and at least one hydrogen bond donor. Isoforms of the CYP2C subfamily are responsible for metabolism of nearly 20% of all small molecule drugs commonly used by humans. The human CYP2C9 gene is highly polymorphic and CYP2C9 single nucleotide polymorphisms (SNPs) have been implicated in clinical drug/drug interactions involving widely-prescribed drugs with narrow therapeutic indices such as warfarin, phenytoin and indomethacin. In addition, for drugs whose pharmacological activity requires metabolism from a pro-drug form (such as the antihypertensive drug losartan), CYP2C9 inhibition or allelic deficiency can lead to decreased drug efficacy. BioVision’s CYP2C9 Activity Assay Kit enables rapid measurement of native or recombinant CYP2C9 activity in biological samples such as liver microsomes. 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. A highly selective irreversible CYP2C9 inhibitor is provided for determination of CYP2C9 activity in heterogeneous biological samples, where other CYP isozymes may contribute to substrate metabolism. The inhibitor displays greater than 100-fold selectivity for CYP2C9 over other CYPs, ensuring targeted inhibition. CYP2C9 specific activity is calculated by running parallel reactions in the presence and absence of the selective inhibitor and subtracting any residual activity detected with the inhibitor present. The kit contains a complete set of reagents sufficient for performing 50 sets of paired reactions (in the presence and absence of inhibitor).
