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Cytochrome P450 2C9 (CYP2C9) Activity Assay Kit (Fluorometric) (CAT#: CB-P240-K) Datasheet

Product Type
Cytochrome P450 2C9 (CYP2C9, EC is a member of the cytochrome P450 monooxidase (CYP) family of microsomal heterologous biological metabolic enzymes. CYP2C9 is mainly expressed in the liver and catalyzes the oxidation of weakly acidic or hydrophobic small molecules containing aromatic moieties and at least one hydrogen bond donor. The isoforms of the CYP2C subfamily are responsible for nearly 20% of the metabolism of all small molecule drugs commonly used in humans. Human CYP2C9 gene is highly polymorphic, and CYP2C9 single nucleotide polymorphism (SNP) has been associated with clinical drug/drug interactions, involving prescription drugs with a wide range of interactions, these drugs have a narrow therapeutic index, such as warfarin, Phenytoin and indomethacin. In addition, for drugs whose pharmacological activity needs to be metabolized from the form of prodrugs (such as the antihypertensive drug Losartan), CYP2C9 inhibition or allele deficiency can lead to reduced drug efficacy. Creative Biolabs' CYP2C9 activity assay kit can quickly measure the activity of natural or recombinant CYP2C9 in biological samples (such as liver microsomes). This assay utilizes a non-fluorescent CYP2C9 substrate, which is converted into a highly fluorescent metabolite detected in the visible light range (Ex / Em = 415/502 nm), thereby ensuring a high signal-to-noise ratio. The interference of autoFluorometric is very small. A highly selective irreversible CYP2C9 inhibitor is provided for the assay of CYP2C9 activity in heterogeneous biological samples, where other CYP isoenzymes may promote substrate metabolism. Compared with other CYPs, the inhibitor has a selectivity of CYP2C9 more than 100 times higher, thus ensuring targeted inhibition. The CYP2C9 specific activity is calculated by performing parallel reactions in the presence and absence of selective inhibitors and subtracting any residual activity detected by the inhibitors present. The kit contains a complete set of reagents, enough to perform 50 paired reactions (in the presence and absence of inhibitors).
100 assays

Figure 1 7-hydroxy-4-(trifluoromethyl) coumarin (7-HFC) standard curve.

Figure 2 Reaction kinetics of fluorogenic substrate metabolism in donor-pooled human liver microsomes.

Figure 3 Specific activity of CYP2C9 in human liver microsome sample.

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