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Cytochrome P450 3A4 (CYP3A4) Activity Assay Kit (Fluorometric)

Highly Sensitive; Includes human CYP3A4 - NADPH Reductase as positive control.
Catalog #: K701
$480.00

Product Details

Cat # +Size K701-200
Size 200 assays
Detection Method Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Eukaryotes
Applications Rapid assessment of native/recombinant CYP3A4 activity in lysates or microsomal fractions prepared from tissues or cells.
Screening of drugs and novel ligands for interaction with native/recombinant CYP3A4.
Features & Benefits • Simple, convenient, highly sensitive
• Fluorescent assay enables determination of CYP3A4, CYP3A5 activity in a variety of biological samples
• The substrate shows low background and a high signal-to-noise ratio
• Kit includes CYP3A inhibitor ketoconazole and a stable, recombinant human CYP3A4 co-expressed with NADPH Reductase as a positive control
Kit Components • CYP3A4 Assay Buffer
• Resorufin Standard (5 mM in DMSO)
• CYP3A4 Inhibitor (Ketoconazole)
• NADPH Generating System (100X)
• β-NADP+ Stock (100X)
• CYP3A4 Substrate
• Recombinant Human CYP3A4
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Cytochrome P450 3A4 (CYP3A4, EC 1.14.13.157) 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 transformed to more hydrophilic products to facilitate excretion from the body. CYP3A4 is expressed in high levels in the liver and intestines, where it catalyzes oxidation of an extraordinarily wide variety of structurally distinct ligands. More than half of all small molecule drugs commonly used by humans are metabolized by CYP3A4. Inhibition of CYP3A4-mediated metabolism is a common cause of adverse drug/drug and drug/food interactions and toxicity. In addition, for drugs whose pharmacological activity requires metabolism from a pro-drug form, CYP3A4 inhibition can lead to decreased drug efficacy. BioVision’s CYP3A4 Activity Assay Kit enables rapid measurement of native or recombinant CYP3A4 activity in biological samples such as liver microsomes. The assay utilizes a non-fluorescent CYP3A4 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 535/587 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. CYP3A4 specific activity is calculated by running parallel reactions in the presence and absence of the potent inhibitor Ketoconazole and subtracting any residual activity detected with the inhibitor present. The kit contains a complete set of reagents sufficient for performing 100 sets of paired reactions (in the presence and absence of inhibitor).


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How does the kit work?
The kit is only designed for studying the effect of different test ligands that can either modulate or inhibit CYP3A4 activity. Therefore, we have a single CYP3A4 specific substrate in the kit. The substrate is metabolized by CYP3A4 to make a fluorogenic end product. The test ligand can either inhibit or activate this process. There could be some cross-reactivity with CYP3A5.
Can K893-100 detect the activity of different isoforms from CYP1A subfamily? or it is highly specific for CYP1A2?
Based on the literature, the substrates should work with rat or mouse CYP enzymes too. K893-100: the substrate can be recognized by both CYP1A1 and CYP1A2.
Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.