Overview

  • 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: Other species, Mammals
  • 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).

  • 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.

  • 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 (10 mg) 1 vial
  • 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 mole of 3-CHC corresponds to the metabolism of one mole of CYP1A2 substrate.

  • 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).

  • CYP1A2 Activity Assay Kit (ab211074) Specific activity of CYP1A2 in pooled human liver microsome sample.

Protocols

References

ab211074 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab211074.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up