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Product Details
| Cat # +Size | K700-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Absorbance (460 nm) |
| Species Reactivity | Eukaryotes |
| Applications | The kit can detect CPR activity in a variety of sample types with a detection limit of ~0.2 mU per well. |
| Features & Benefits | • Only 96-well plate format CPR assay kit on the market • Only kit to use a synthetic colorimetric probe and a NADPH generating system • Rapid, convenient and sensitive • Kit contains all necessary reagents for accurate measurement of CPR activity down to 0.2 mU per well in a wide variety of sample types |
| Kit Components | • CPR Assay Buffer • G6P Standard • Inhibitor (Diphenyleneiodonium Chloride, 10 mM) • NADPH Substrate Mix • G6P Standard Developer • Human CPR Positive Control |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
NADPH-cytochrome P450 reductase (CPR, EC 1.6.2.4) is a ~78 kDa membrane-bound flavoenzyme that catalyzes the transfer of electrons from NADPH to members of the cytochrome P450 monooxidase (CYP) enzyme family in the endoplasmic reticulum. CPR contains two tightly bound flavin cofactors, FAD and FMN, which participate in the sequential transfer of electrons from NADPH→FAD→FMN→CYP, oxidizing NADPH to NADP+ and reducing the CYP heme moiety to the substrate- and oxygen-binding ferrous state. As CPR is required for the function of all CYP isozymes, it plays a critical role in the metabolism of drugs, organic pollutants and other xenobiotic compounds, in addition to its role in biosynthesis of certain vitamins and steroid hormones. BioVision’s cytochrome P450 reductase activity assay kit couples oxidation of NADPH by CPR to reduction of a nearly colorless probe into a brightly colored product with an absorbance peak at 460 nm, with the rate of color generation being directly proportional to CPR activity. The NADPH utilized by CPR is generated in situ from β-NADP+ via oxidation of glucose-6-phosphate (G6P) to 6-phospho-D-glucono-1,5-lactone by glucose-6-phosphatase dehydrogenase (G6PDH). The kit can be used to determine CPR activity in a variety of samples, with a detection limit of ~0.2 mU of CPR activity per reaction. For assessment of CPR activity in crude biological samples that may have extraneous reductases capable of reducing the substrate, an inhibitor of NADPH-dependent flavoproteins is included. In this case, the specific CPR activity may be calculated by running parallel reactions in the presence and absence of inhibitor and subtracting any residual activity detected with the inhibitor present. The kit contains sufficient reagents for performing 100 reactions in a 96-well plate format.
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Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Yuan Zheng, Lysosomal Proteases Are a Determinant of Coronavirus Tropism. J Virol, nov 2018; 30258004.
For more citations of this product click here
