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Cytochrome P450 Reductase (CPR) Activity Kit (Colorimetric) (CAT#: CB-P246-K) Datasheet

Product Type
Kit
Description
NADPH-Cytochrome P450 Reductase (CPR, EC 1.6.2.4) is a 78 kDa membrane-bound flavanase that catalyzes the transfer of electrons from NADPH to the cytochrome P450 monooxidase (CYP) enzyme family in the endoplasmic reticulum in. CPR contains two tightly bound flavin cofactors FAD and FMN, which participate in the sequential transfer of electrons from NADPHADMNYP, oxidize NADPH to NADP +, and partially reduce CYP heme to the ferrous state that is combined with the substrate and oxygen. Since CPR is necessary for the functions of all CYP isoenzymes, it not only plays a role in the biosynthesis of certain vitamins and steroid hormones, but also plays a role in the metabolism of drugs, organic pollutants and other heterologous compounds. play a key role in. Creative Biolabs' cytochrome P450 reductase activity assay kit can reduce the oxidation of CAD on NADPH and the almost colorless probe into a brightly colored product with an absorption peak at 460 nm, and the color development rate is proportional to the CPR activity. Proportional. The NADPH used in CPR is derived from Beta-NADP+ oxidizing glucose-6-phosphate (G6P) to 6-phosphate-D-glucosyl-1,5-lactone through glucose-6-phosphatase dehydrogenase (G6PDH) Generated in situ. The kit can be used to determine CPR activity in various samples, and the assay limit of CPR activity for each reaction is about 0.2 mU. In order to evaluate the CPR activity in crude biological samples, these crude samples may have exogenous reductase capable of reducing the substrate, and therefore contain NADPH-dependent flavoprotein inhibitors. In this case, the CPR specific activity can be calculated by performing parallel reactions in the presence and absence of inhibitors and subtracting any residual activity detected by the inhibitors present. The kit contains enough reagents to perform 100 reactions in a 96-well plate format.
Size
100 assays

Figure 1 G6P Standard curve.

Figure 2 Reaction kinetics of recombinant human CPR (positive control) and rat microsomal CPR (with and without inhibitor).

Figure 3 Relative CPR activity detected in rat liver microsomes (RLM, 25 µg total protein) and HepG2 cell lysate (40 µg total protein).

All products and services are for Research Use Only. Do Not use in humans.

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