CYP1A2 Activity Assay Kit (Fluorometric) (ab211074)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Cell Lysate, Microsomes, Tissue Lysate
Overview
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Product name
CYP1A2 Activity Assay Kit (Fluorometric)
See all Cytochrome P450 1A2 kits -
Detection method
Fluorescent -
Sample type
Cell Lysate, Microsomes, Tissue Lysate -
Assay type
Enzyme activity (quantitative) -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
CYP1A2 Activity Assay Kit (Fluorometric) (ab211074) allows rapid measurement of native or recombinant cytochrome P450 1A2 (CYP1A2) activity in biological samples such as liver microsomes. The assay utilizes a non-fluorescent CYP1A2 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 406/468 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. A selective CYP1A2 inhibitor is provided for determination of CYP1A2 activity in heterogeneous biological samples, where other CYP isozymes may contribute to substrate metabolism. The inhibitor displays greater than 20-fold selectivity for CYP1A2 over other CYPs, ensuring targeted inhibition. CYP1A2 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).
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Notes
Cytochrome P450 1A2 (CYP1A2, EC 1.14.14.1) is a member of the cytochrome P450 monooxidase (CYP) family of microsomal xenobiotic metabolism enzymes. CYPs are membrane-bound hemeproteins responsible for Phase I biotransformation reactions, in which lipophilic drugs and other xenobiotic compounds are converted to more hydrophilic products to facilitate excretion from the body. CYP1A2 is primarily expressed in liver, intestinal and olfactory mucosal tissue and catalyzes oxidation of polyaromatic and heterocyclic molecules such as aromatic amines. CYP1A2 is responsible for metabolism of approximately 10% of all small molecule drugs commonly used by humans. Polymorphisms in the human CYP1A2 gene have been implicated in clinical drug/drug interactions involving widely-used drugs, including caffeine, theophylline and the antipsychotic clozapine. Isoforms of the CYP1A subfamily are also involved in metabolic activation of environmental pro-carcinogens present in cigarette smoke and combustion exhaust fumes.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests β-NADP+ Stock 100X 1 vial 3-CHC Standard 1 vial CYP1A2 Assay Buffer 1 x 100ml CYP1A2 Inhibitor (α-naphthoflavone) 1 vial CYP1A2 Substrate 1 vial NADPH Generating System 100X 1 vial Recombinant Human CYP1A2 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. Most active in catalyzing 2-hydroxylation. Caffeine is metabolized primarily by cytochrome CYP1A2 in the liver through an initial N3-demethylation. Also acts in the metabolism of aflatoxin B1 and acetaminophen. Participates in the bioactivation of carcinogenic aromatic and heterocyclic amines. Catalizes the N-hydroxylation of heterocyclic amines and the O-deethylation of phenacetin. -
Tissue specificity
Liver. -
Sequence similarities
Belongs to the cytochrome P450 family. -
Cellular localization
Endoplasmic reticulum membrane. Microsome membrane. - Information by UniProt
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Alternative names
- Aryl hydrocarbon hydroxylase
- CP 12
- CP12
see all
Images
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CYP1A2 Activity Assay Kit (Fluorometric) (ab211074)
Typical 3-CHC standard calibration curve. One mole of 3-CHC corresponds to the metabolism of one mole of CYP1A2 substrate.
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CYP1A2 Activity Assay Kit (Fluorometric) (ab211074)CYP1A2 Activity Assay Kit (ab211074) Reaction kinetics of fluorogenic substrate metabolism in donor-pooled human liver microsomes (0.25 mg/mL) at 37°C in the presence and absence of the CYP1A2 inhibitor α-naphthoflavone (the solvent control contained assay buffer with 0.4% acetonitrile).
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CYP1A2 Activity Assay Kit (Fluorometric) (ab211074)CYP1A2 Activity Assay Kit (ab211074) Specific activity of CYP1A2 in pooled human liver microsome sample.
