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Cytochrome P450 1A2 (CYP1A2) Activity Assay Kit (Fluorometric) (CAT#: CB-P236-K) Datasheet

Product Type
Cytochrome P450 1A2 (CYP1A2, EC is a member of the cytochrome P450 monooxidase (CYP) family of microsomal heterologous biological metabolic enzymes. CYP is the membrane-bound hemoglobin responsible for the phase I biotransformation reaction, in which lipophilic drugs and other heterogeneous biological compounds are converted into more hydrophilic products to promote human excretion. CYP1A2 is mainly expressed in liver, intestine and olfactory mucosa tissues, and catalyzes the oxidation of polycyclic aromatic and heterocyclic molecules (such as aromatic amines). CYP1A2 is responsible for about 10% of the metabolism of all small molecule drugs commonly used in humans. The polymorphism of the human CYP1A2 gene is related to clinical drug/drug interactions involving widely used drugs (including caffeine, theophylline and the antipsychotic clozapine). Subtypes of the CYP1A subfamily are also involved in the metabolic activation of environmental carcinogens in cigarette smoke and combustion exhaust gas. Creative Biolabs' CYP1A2 activity assay kit can quickly measure the activity of natural or recombinant CYP1A2 in biological samples (such as liver microsomes). The assay utilizes a non-fluorescent CYP1A2 substrate, which is converted into a highly fluorescent metabolite detected in the visible light range (Ex / Em = 406/468 nm), thus ensuring a high signal-to-noise ratio. The interference of autoFluorometric is very small. A selective CYP1A2 inhibitor is provided to determine the CYP1A2 activity in heterogeneous biological samples, in which other CYP isoenzymes may promote substrate metabolism. The inhibitor's selectivity for CYP1A2 is more than 20 times that of other CYPs, thereby ensuring targeted inhibition. The specific activity of CYP1A2 is calculated by performing parallel reactions in the presence and absence of the selective inhibitor and subtracting any residual activity found in the presence of the inhibitor. 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 3-cyano-7-hydroxycoumarin (3-CHC) standard curve.

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

Figure 3 Specific activity of CYP1A2 in human liver microsome sample.

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