Overview

  • Product name

    CYP1A2 Inhibitor Assay Kit (Fluorometric)
    See all Cytochrome P450 1A2 kits
  • Detection method

    Fluorescent
  • Sample type

    Inhibitor compounds
  • Assay type

    Quantitative
  • Product overview

    CYP1A2 Inhibitor Screening Kit (Fluorometric) (ab211075) allows rapid screening of drugs and other new chemical entities (NCEs) for compound-CYP1A2 interaction in a reliable, high-throughput fluorescence-based assay. The kit provides a yeast microsomal preparation of human CYP1A2 and human cytochrome P450 reductase (CPR) enzymes. 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.  The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96 well plate format.

  • 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 planar 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 methylxanthines (caffeine and theophylline), ciprofloxacin and a number of antidepressants and antipsychotics. Isoforms of the CYP1A subfamily are also involved in metabolic activation of environmental pro-carcinogens in cigarette smoke and combustion exhaust fumes.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    β-NADP+ Stock 100X (2 mg) 1 vial
    3-CHC Standard (2 mg) 1 vial
    CYP1A2 Assay Buffer 1 x 100ml
    CYP1A2 Inhibitor (α-naphthoflavone) (2 mg) 1 vial
    CYP1A2 Substrate (2 mg) 1 vial
    NADPH Generating System 100X (5 mg) 1 vial
    Recombinant Human CYP1A2 (2 x 5 mg) 2 vials
  • Research areas

  • 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
  • Alternative names

    • Aryl hydrocarbon hydroxylase
    • CP 12
    • CP12
    • CP1A2_HUMAN
    • CYP1A2
    • CYPIA2
    • Cytochrome P(3)450
    • Cytochrome P450 1A2
    • Cytochrome P450 4
    • Cytochrome P450 family 1 polypeptide 2
    • Cytochrome P450 family 1 subfamily A polypeptide 2
    • Cytochrome P450 subfamily I aromatic compound inducible polypeptide 2
    • Cytochrome P450-P3
    • Cytochrome P4501A2
    • Dioxin inducable P3 450
    • Flavoprotein linked monooxygenase
    • Flavoprotein-linked monooxygenase
    • Microsomal monooxygenase
    • P(3)450
    • P3 450
    • P450 4
    • P450 form 4
    • P450 P3
    • P450(PA)
    • Xenobiotic monooxygenase
    see all

Images

  • Typical 3-CHC standard calibration curve. One mol of 3-CHC corresponds to the metabolism of one mol of CYP1A2 substrate.

  • Reaction kinetics of recombinant human CYP1A2 enzyme at 37°C in the presence and absence of the indicated inhibitor (solvent control reaction includes final concentration of 0.6% acetonitrile).

  • Dose-response curves for various CYP1A2 ligands of differing structural and mechanistic classes: the competitive CYP1A2 inhibitor α-naphthoflavone, the antidepressant fluvoxamine, the tricyclic antipsychotic chlorpromazine and the endogenous neurohormone melatonin (a CYP1A2 substrate). For dose-response curves, percent activity was calculated for each concentration of inhibitor by comparison to activity of reactions containing no inhibitor. For each CYP1A2 inhibitor, IC50 values were derived by 4-parameter logistic curve fitting with each point representing the mean ± SEM of at least three replicates. Assays were performed according to the kit protocol.

     

Protocols

References

ab211075 has not yet been referenced specifically in any publications.

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