Cytochrome P450 1A2 (CYP1A2) Inhibitor Screening Kit (Fluorometric) (BN01068)

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BN01068
  • Cytochrome P450 1A2 (CYP1A2) Inhibitor Screening Kit (Fluorometric) (BN01068)
  • Cytochrome P450 1A2 (CYP1A2) Inhibitor Screening Kit (Fluorometric) (BN01068)
  • Cytochrome P450 1A2 (CYP1A2) Inhibitor Screening Kit (Fluorometric) (BN01068)
€535

Description

ELISA Kit Technical ManualMSDS

Cytochrome P450 1A2 (CYP1A2) Inhibitor Screening Kit (Fluorometric)

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. Assay Genie's CYP1A2 Inhibitor Screening Kit enables 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.

Figure: (a) 3-cyano-7-hydroxycoumarin (3-CHC) standard curve. One mole of 3-CHC corresponds to the metabolism of one mole of CYP1A2 substrate. (b) Reaction kinetics of recombinant human CYP1A2 enzyme at 37°C in the presence and absence of the indicated CYP1A2 inhibitors (the solvent control reaction contained assay buffer with 0.6% acetonitrile). (c) 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, IC 50 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.

Key Information Description

Product SKU

BN01068

Size

100 assays

Detection Method

Fluorescence (Ex/Em 406/468 nm)

Species Reactivity

Eukaryotes

Applications

  • Rapid, high-throughput screening of drugs and novel ligands.
  • Development of structure-activity relationship (SAR) models to predict CYP1A2 inhibition liability of novel compounds.
  • Prediction of adverse drug-drug interaction potential and bioavailability for compounds metabolized by CYP1A2.

Features and Benefits

  • Simple, convenient, highly sensitive
  • Fluorescent assay enables easy determination of CYP1A2 activity in a variety of biological samples
  • The substrate shows low background and a high signal-to-noise ratio
  • Kit includes CYP1A2 inhibitor (naphthoflavone) and a stable, recombinant human CYP1A2 co-expressed with NADPH
  • Reductase as a positive control

Kit Components

  • CYP1A2 Assay Buffer
  • 3-CHC Standard
  • CYP1A2 Inhibitor (naphthoflavone)
  • NADPH Generating System (100X)
  • Beta-NADP+ Stock (100X)
  • CYP1A2 Substrate
  • Recombinant Human CYP1A2

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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