Cytochrome P450 2C9 (CYP2C9) Activity Assay Kit (Fluorometric) (ab273336)
Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate
- Sample type: Cell Lysate, Microsomes, Purified protein, Tissue Lysate
Overview
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Product name
Cytochrome P450 2C9 (CYP2C9) Activity Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Cell Lysate, Microsomes, Purified protein, Tissue Lysate -
Assay type
Enzyme activity -
Assay duration
Multiple steps standard assay -
Product overview
Cytochrome P450 2C9 (CYP2C9) Activity Assay Kit (Fluorometric) (ab273336) enables the rapid measurement of native or recombinant CYP2C9 activity in biological samples such as liver microsomes.
The assay utilizes a non-fluorescent CYP2C9 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 415/502 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. A highly selective irreversible CYP2C9 inhibitor is provided for determination of CYP2C9 activity in heterogeneous biological samples, where other CYP isozymes may contribute to substrate metabolism. The inhibitor displays greater than 100-fold selectivity for CYP2C9 over other CYPs, ensuring targeted inhibition. CYP2C9 specific activity is calculated by running parallel reactions in the presence and absence of the selective inhibitor and subtracting any residual activity detected with the inhibitor present.
The kit contains a complete set of reagents sufficient for performing 50 sets of paired reactions (in the presence and absence of inhibitor). -
Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Beta-NADP+ Stock (100X) 1 vial 7-HFC Standard Lyophilized 1 vial CYP2C9 Assay Buffer 1 x 100ml CYP2C9 Inhibitor (Tienilic Acid) Lyophilized 1 vial CYP2C9 Substrate Lyophilized 1 vial NADPH Generating System (100X) Lyophilized 1 vial Recombinant Human CYP2C9 Lyophilized 1 vial -
Research areas
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Function
Cytochromes P450 are a group of heme-thiolate monooxygenases. In liver microsomes, this enzyme is involved in an NADPH-dependent electron transport pathway. It oxidizes a variety of structurally unrelated compounds, including steroids, fatty acids, and xenobiotics. This enzyme contributes to the wide pharmacokinetics variability of the metabolism of drugs such as S-warfarin, diclofenac, phenytoin, tolbutamide and losartan. -
Sequence similarities
Belongs to the cytochrome P450 family. -
Cellular localization
Endoplasmic reticulum membrane. Microsome membrane. - Information by UniProt
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Alternative names
- (R)-limonene 6-monooxygenase
- (S)-limonene 6-monooxygenase
- (S)-limonene 7-monooxygenase
see all
Images
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Example data
Example data.
7-hydroxy-4-(trifluoromethyl) coumarin (7-HFC) standard curve. One mole of 7-HFC corresponds to the metabolism of one mole of CYP2C9 substrate.
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Example dataExample data.
Reaction kinetics of fluorogenic substrate metabolism in donor-pooled human liver microsomes (0.125 mg/mL) at 37°C in the presence and absence of the CYP2C9 inhibitor tienilic acid (the solvent control contained assay buffer with 0.3% acetonitrile).
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Example dataExample data.
Specific activity of CYP2C9 in human liver microsome sample.