The assay is optimized and validated at 37 °C. Running the assay at 25 °C may slow the enzyme reaction and reduce the absorbance change at 340 nm. Results may therefore differ from those obtained using the standard protocol, and independent optimization would be required. Compatibility with dimethyl sulfoxide has not been established. Use the lowest possible final dimethyl sulfoxide concentration, keep the concentration consistent across all samples and vehicle controls, and include a control without dimethyl sulfoxide to assess the effect on enzyme activity.
CS1090
HMG-CoA Reductase Assay Kit
sufficient for 30 assays (1 mL), sufficient for 100 assays (200 μL)
Sinónimos:
HMGR Assay Kit
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| Talla/SKU | Disponibilidad | Precio |
|---|---|---|
1 kit | Póngase en contacto con nuestro Servicio de Atención al Cliente para disponibilidad | 1040,00 € |
Acerca de este artículo
NACRES:
NA.28
UNSPSC Code:
12161503
eCl@ss:
32160702
1040,00 €
Póngase en contacto con nuestro Servicio de Atención al Cliente para disponibilidad
Servicio técnico
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Permítanos ayudarleQuality Segment
usage
sufficient for 100 assays (200 μL), sufficient for 30 assays (1 mL)
shipped in
dry ice
storage temp.
−70°C
Gene Information
human ... HMGCR(3156)
General description
The HMG-CoA Reductase Assay Kit is an important tool for the basic research of cholesterol and other related metabolic pathways.
Application
The HMGR activity kit is designed for the detection of HMG-CoA reductase activity. A major function of the kit is to screen for various inhibitors and activators of the purified catalytic subunit of the enzyme, HMG-CoA reductase, which may play a crucial role in therapeutics. The assay is based on the spectrophotometric measurement of the decrease in absorbance at 340 nm, which represents the oxidation of NADPH by the catalytic subunit of HMGR in the presence of the substrate HMG-CoA.
Biochem/physiol Actions
HMGR (3-hydroxy-3-methylglutaryl-CoA reductase) is a transmembrane glycoprotein located on the endoplasmic reticulum. This enzyme catalyzes the four-electron reduction of HMG-CoA to coenzyme A (CoA) and mevalonate, which is the rate-limiting step in sterol biosynthesis. The activity of HMGR is controlled through synthesis, degradation, and phosphorylation in order to maintain the concentration of mevalonate derived products. In addition to the physiological regulation of HMGR, the human enzyme has been targeted successfully by drugs in the clinical treatment of high serum cholesterol levels. Controlling serum cholesterol levels has an important therapeutic role since hypercholesterolemia often leads to the development of atherosclerosis and subsequently to cardiovascular pathologies, which might result in myocardial infarction and stroke. It has been suggested that a disturbance of cholesterol homeostasis contributes to the development of a chronic inflammatory state.
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Este artículo | |||
|---|---|---|---|
| description sufficient for 30 assays (1 mL), sufficient for 100 assays (200 μL) | description solution | description clone IgG-A9, from mouse | description from rabbit, purified by affinity chromatography |
| Gene Information human ... HMGCR(3156) | Gene Information human ... HMGCR(3156) | Gene Information human ... HMGCR(3156) | Gene Information human ... HMGCR(3156) |
| Quality Level 200 | Quality Level 200 | Quality Level 100 | Quality Level 100 |
| usage sufficient for 30 assays (1 mL), sufficient for 100 assays (200 μL) | usage - | usage - | usage - |
| shipped in dry ice | shipped in dry ice | shipped in ambient | shipped in wet ice |
| storage temp. −70°C | storage temp. −70°C | storage temp. - | storage temp. - |
Clase de almacenamiento
10 - Combustible liquids
Palabra de advertencia
Warning
Códigos de peligro
Indicaciones de precaución
Hazard Classifications
Eye Irrit. 2 - STOT SE 2
Órganos diana
Eyes,Central nervous system
WGK
WGK 3
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Hello there, here we don't have spectrophotometer with heating element. Is it possible to do the assay at around room temperature (25 Celsius)? Another thing, how is the DMSO effect toward the enzyme?
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How can I determine the shelf life / expiration / retest date of this product?
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If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/418/501/product-dating-information-06-25-mk.pdf¿Le ha resultado útil?
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How many samples can we analyse using 1Kt?
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Tables 1 and 2 provide specific instructions for sample preparation for reaction volumes in a cuvette (1 mL) or a 96 well plate. The limiting reagent based upon the supplied kit components is the HMG-COA. There is 2 mL of HMG-COA supplied and 0.18 mL used in total for the blank, activity and inhibitor well preparations. This means that approximately 33 wells or 30 assays can be performed as indicated in the kit bulletin.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/240/783/cs1090pis-ms.pdf
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How is shipping temperature determined? And how is it related to the product storage temperature?
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Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf
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What is the concentration of the HMG CoA substrate (S7447)?
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The concentration of the HMG CoA substrate, product number S7447, is 0.5% in water.
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Is this suitable for the analysis of compounds dissolved in DMSO for comparing the ability of some compounds to inhibit HMGCR? Specifically, using 1 ul of the compound solution (0.05, 0.5, 5 mM stock buffer) and 1 ul DMSO (solvent alone) as control, the signal @ 340 nM did not significantly decrease after a 10-minute kinetic program (signal capture every 20 seconds according to the protocol). There was only a 15-20 percent decrease in the read. Is this normal, or is DMSO incompatible with the enzyme reaction, or can some improvement be made?
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As per the QC specialist group, in the product CS1090, there isn't any experience with DMSO (all kit's components are without DMSO), so the information cannot provided about DMSO incompatibility. After reviewing the raw data, a good slope was observed with the enzyme; for example, with 6ug (10ul) enzyme, the slope was 0.3-0.5.
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•What is the concentration of the HMG-CoA (Cat.No. S7447)? and What solvent is it dissolved in?
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The concentration of HMG-CoA is the composition of 99.5 % water and 0.5 % (DL-3-Hydroxy-3-methylglutaryl) coenzyme aso-salt trihydrate.
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Product CS1090, kit component of HMG-CoA Reductase Assay Kit Product I5909, Pravastatin, is the concentration 100uM correct?
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Yes, the concentration for Pravastatin in this kit is 100 micromolar (100 μM)
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What is the lowest concentration that you can use Pravastatin and observe inhibition in Product CS1090, HMG-CoA Reductase Assay kit?
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For Product CS1090, HMG-CoA Reductase Assay kit we do not observe any inhibition when using Pravastatin at 25 or 50 nM. We observe inhibition of HMG-CoA Reductase with 250 nM pravastatin.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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