Cytochrome P450 Reductase Activity Assay试剂盒(Colorimetric) (ab204704)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell Lysate, Microsomes, Purified protein, Tissue Lysate
- Sensitivity: 0.2 mU/well
概述
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产品名称
Cytochrome P450 Reductase Activity Assay试剂盒(Colorimetric)
参阅全部 Cytochrome P450 Reductase 试剂盒 -
检测方法
Colorimetric -
样品类型
Cell Lysate, Microsomes, Purified protein, Tissue Lysate -
检测类型
Enzyme activity -
灵敏度
0.2 mU/well -
种属反应性
与反应: Mammals, Other species -
产品概述
Cytochrome P450 Reductase Activity Assay Kit (Colorimetric) (ab204704) couples the oxidation of NADPH by cytochrome P450 reductase (CPR) to the reduction of a nearly colorless probe into a brightly colored product with an absorbance peak at OD=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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说明
This product is manufactured by BioVision, an Abcam company and was previously called K700 Cytochrome P450 Reductase (CPR) Activity Kit (Colorimetric). K700-100 is the same size as the 100 test size of ab204704.
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.
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平台
Microplate reader
性能
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存放说明
Store at -20°C. Please refer to protocols. -
组件 100 tests CPR Assay Buffer 1 x 50ml CPR Inhibitor 1 x 100µl CPR Standard (G6P) 1 vial G6P Standard Developer 1 vial Human CPR Positive Control 1 vial NADPH Substrate Mix 1 vial -
研究领域
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功能
This enzyme is required for electron transfer from NADP to cytochrome P450 in microsomes. It can also provide electron transfer to heme oxygenase and cytochrome B5. -
疾病相关
Defects in POR are the cause of adrenal hyperplasia variant type (AHV) [MIM:201750]; also known as Antley-Bixler syndrome-like phenotype with disordered steroidogenesis. AHV is a rare variant of congenital adrenal hyperplasia. It is an autosomal recessive disorder with apparent combined P450C17 and P450C21 deficiency. Affected girls are born with ambiguous genitalia, but their circulating androgens are low and virilization does not progress. Conversely, affected boys are sometimes born undermasculinized. Boys and girls can also present with bone malformations, in some cases resembling the pattern seen in patients with Antley-Bixler syndrome.
Defects in POR are a cause of isolated disordered steroidogenesis (IDS) [MIM:201750]. -
序列相似性
In the C-terminal section; belongs to the flavoprotein pyridine nucleotide cytochrome reductase family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. -
细胞定位
Endoplasmic reticulum membrane. Anchored to the ER membrane by its N-terminal hydrophobic region. - Information by UniProt
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别名
- CPR
- CYPOR
- Cytochrome p450 oxidoreductase
see all
图片
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Typical G6P Standard curve using Cytochrome P450 Reductase Activity Assay Kit (Colorimetric) (ab204704). One mole G6P corresponds to one mole of β-NADP+ reduced to NADPH, which subsequently generates one mole of reduced substrate.
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Reaction kinetics of recombinant human CPR (positive control) and rat microsomal CPR (with and without inhibitor).
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Relative CPR activity detected in rat liver microsomes (RLM, 25 µg total protein) and HepG2 cell lysate (40 µg total protein).
数据表及文件
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SDS download
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Datasheet download
文献 (1)
ab204704 被引用在 1 文献中.
- Zlotnik D et al. P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB. Stem Cell Reports 17:2050-2063 (2022). PubMed: 35961311