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GRSA Sigma

Glutathione Reductase Assay Kit

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Properties

Related Categories Cell Analysis, Cell Biology, Cell Signaling and Neuroscience, Cell Stress, Nitric Oxide and Cell Stress,
usage   sufficient for 100 tests
storage temp.   2-8°C
Gene Information   human ... GSR(2936)

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Analysis Note

UV assay: One unit will cause the oxidation of 1.0 μmole of NADPH at 25 °C at pH 7.5.
Colorimetric assay: One unit will cause the reduction of 1.0 μmole of DTNB to TNB at 25 °C at pH 7.5.

Biochem/physiol Actions

Glutathione reductase (EC 1.6.4.2) (GR) is a ubiquitous enzyme that catalyzes the reduction of oxidized glutathione (GSSG) to glutathione (GSH). Glutathione reductase is essential for the glutathione redox cycle that maintains adequate levels of reduced cellular GSH, which serves as an antioxidant reacting with free radicals and organic peroxides. Glutathione is also a substrate for the glutathione peroxidases and glutathione S-transferases in the detoxification of organic peroxides and the metabolism of xenobiotics. This kit contains reagents for the spectrophotometric determination of glutathione reductase activity either by following the decrease in absorbance caused by the oxidation of NADPH at 340 nm (UV assay) or the increase in absorption caused by the reduction of dithiobis(2-nitrobenzoic acid) (DTNB) at 412 nm (colorimetric assay). This kit provides reagents for a spectrophotometric assay for measuring the activity of glutathione reductase either by following the decrease in A340 caused by the oxidation of NADPH or the increase in A412 caused by the reduction of dithiobis(2-nitrobenzoic acid).

This kit provides reagents for a spectrophotometric assay for measuring the activity of glutathione reductase either by following the decrease in A340 caused by the oxidation of NADPH or the increase in A412 caused by the reduction of dithiobis(2-nitrobenzoic acid).

Price and Availability


All labs need water

Kit component only

Description

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Glutathione Reductase Assay Buffer 125 mL SDS    
Glutathione Reductase Dilution Buffer 100 mL SDS    
Glutathione Reductase Positive Control 1 mL/vial SDS    

Standard component

Description

Product #

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5,5′-Dithiobis(2-nitrobenzoic acid) 50 mg SDS D8130
Glutathione, oxidized, disodium salt 100 mg SDS G4626
β-Nicotinamide adenine dinucleotide 25 mg SDS N6505
Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

Certificate of Origin

Frequently Asked Questions

How does the storage temperature relate to shipping conditions?
The storage conditions that a Sigma-Aldrich catalog and label recommend for products are deliberately conservative. For many products, long-term storage at low temperatures will increase the time during which they are expected to remain in specification and therefore are labeled accordingly. Where short-term storage, shipping time frame, or exposure to conditions other than those recommended for long-term storage will not affect product quality, Sigma-Aldrich will ship at ambient temperature. The products sensitive to short-term exposure to conditions other than their recommended long-term storage are shipped on wet or dry ice. Ambient temperature shipping helps to control shipping costs for our customers. At any time, our customers can request wet- or dry-ice shipment, but the special handling is at customer expense if our product history indicates that the product is stable for regular shipment. http://www.sigmaaldrich.com/etc/medialib/docs/Sigma-Aldrich/General_Information/shippingandlong-termstoragev3.Par.0001.File.tmp/shippingandlong-termstoragev3.pdf
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
In your product bulletin you say you have tested Product GRSA, Glutathione Reductase Assay Kit, on rabbit reticulocyte lysate and Saccharomyces extract. Have you tested it on other species?
At this time, we have used the kit to test only these species. We have received inquiries concerning its use for rat and higher plant tissue, but we have no data at this time.
Do you know if Product GRSA, Glutathione Reductase Assay Kit, can be used with a 96 well plate reader?
We have used this kit ony with a UV/Vis spectrophotometer with 1 cm pathlength cuvettes. We have not attempted to modify it for use with a plate reader.
Is Product GRSA, Glutathione Reductase Assay Kit, compatible with your CelLytic cell lysis reagents?
We have not tried using this kit with our CelLytic reagents. We believe it should be suitable for use with C2978 for mammalian cell lysis or C4882 for yeast cell lysis.
Is Product GRSA, Glutathione Reductase Assay Kit, suitable for use for GR determination in whole blood?
We do not recommend the kit for use with whole blood due to interference from hemoglobin.
In your brochure for Nitric Oxide and Cell Stress you present absorbance data from yeast lysis. How were the yeast prepared?
Here is the procedure as recommended by our Production group (March 2008): Grow S. cerevisiae (we used GP460 private strain) on YEPD medium; harvest cells by centrifugation. Wash the cells [7g]  with 20 ml of 20 mM HEPES buffer, pH 7.0. Discard supernatant. Add 1 ml HEPES buffer to make a thick slurry and pour dropwise into liquid nitrogen. Take approx. 2 g of frozen pellet and homogenize with UltraTurrax T-25 while still in liquid nitrogen to make a fine powder. After all the liquid nitrogen has evaporated, take approximately 2 mL of HEPES buffer and add to the 2 g of finely powdered cells. Centrifuge for 5 min in an Eppendorf (benchtop) centrifuge at maximum speed. The supernatant  liquid is the source of the enzyme. The total amount of protein received from this procedure was 21 mg (8.8 mg/mL in 2.4 mL total volume). We determined an activity of approximately 0.3 units/mL, using both the UV and DTNB assays.
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Protocols & Articles
Peer-Reviewed Papers
15

References

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Description

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