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HMG-CoA Reductase Activity/Inhibitor Screening Kit (Colorimetric)

Simple assay to measure activity of purified enzyme and screening inhibitors/activators of HMGR
Catalog #: K588

In stock

$595.00

Product Details

Cat # +Size K588-100
Size 100 assays
Detection Method Absorbance (OD 340 nm)
Applications Measurement of HMG-CoA reductase activity
Screening/studying/characterizing HMG-CoA reductase inhibitors/activators
Features & Benefits • Simple, rapid & convenient
• Limit of detection: below 0.05 mU
• Includes inhibitor (Atorvastatin)
Kit Components • HMG-CoA Reductase Assay Buffer
• HMG-CoA Reductase
• HMG-CoA
• NADPH
• Inhibitor (Atorvastatin, 10 mM)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

HMG-CoA reductase (3-hydroxy-3-methyl-glutaryl-CoA reductase or HMGR) (EC 1.1.1.34) is the rate-controlling enzyme of the mevalonate pathway, the metabolic pathway that produces cholesterol from acetyl-CoA. In an NADPH-dependent reaction, HMG-CoA reductase reduces HMG-CoA to generate mevalonate and CoA. The enzyme is target of a group of cholesterol-lowering drugs known as statins. Inhibition of HMG-CoA reductase induces expression of LDL receptors in the liver, which lowers plasma concentration of cholesterol. BioVision’s HMG-CoA Reductase Activity Assay Kit measures utilization of NADPH, which can be measured by the decrease of absorbance (OD 340 nm). The kit is suitable for measuring activity of purified enzyme and screening inhibitors/activators of HMG-CoA reductase. The limit of detection is below 0.05 mU.


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Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
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 yopur regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
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.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Is the enzyme included in the kit the full enzyme, or only catalytic site?
The enzyme in the kit is HMG-CoA Reductase Catalytic Domain (GST Tagged), human recombinant, Biovision # 7841. The enzyme has a molecular weight of 75.9 kDa (426–888 aa, NT GST Tag).
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
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