High Throughput Glutathione Reductase Assay
High Throughput Glutathione Reductase Assay SummaryDesigned to measure cellular levels of glutathione reductase using a 96-well plate.
• Suitable for cell lysates, erythrocyte lysates, and tissue homogenate
• Sufficient reagents for 500 assays
• 96 well format for high throughput screening
Why Use the Glutathione Reductase Assay Kit?
The Glutathione Reductase Assay Kit is a spectrophotometric assay in which the oxidation of NADPH to NADP+ is monitored by the decrease in absorbance at 340 nm. This rate of decrease in absorbance at 340 nm is directly proportional to the glutathione reductase activity in the sample because the enzyme is present at rate limiting concentrations. This kit provides the user with all the reagents and plates to easily and rapidly assay for glutathione reductase in cell and tissue extracts.
• Glutathione Reductase (1U/mL)
• 20% Triton X-100
• 96-Well Plate
• 10X GR Buffer
• GSSG Solution
For research use only. Not for diagnostic use.
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Citations for High Throughput Glutathione Reductase Assay
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Citations: Showing 1 - 3
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Altered glutathione homeostasis in heart augments cardiac lipotoxicity associated with diet-induced obesity in mice.
Authors: Ghosh S, Sulistyoningrum D, Glier M, Verchere C, Devlin A
J Biol Chem, 0;286(49):42483-93. 0
Amount and source of dietary copper affects small intestine morphology, duodenal lipid peroxidation, hepatic oxidative stress,and mRNA expression of hepatic copper regulatory proteins in weanling pigs.
Authors: Fry R, Ashwell M, Lloyd K, O'Nan A, Flowers W, Stewart K, Spears J
J Anim Sci, 0;90(9):3112-9. 0
Mitochondrial oxidative stress-induced epigenetic modifications in pancreatic epithelial cells.
Authors: Mishra P, Raghuram G, Jain D, Jain S, Khare N, Pathak N
Int J Toxicol, 0;33(2):116-29. 0
Is this kit capable of measuring serum/plasma samples?
This kit may be used to measure Glutathione Reductase activity in serum or plasma samples within the detection range of the assay.
What method is recommended to remove hemoglobin from red blood cell lysate?
Over 95% of the hemoglobin can be removed by adding Zinc Sulfate to a final concentration of 500 uM and centrifuging to remove the precipitate. For more information, please see: Sajan MP, and Kulkarni AP. "A simple and rapid method for hemoglobin removal from mammalian tissue cytosol by zinc sulfate and its application to the study of Glutathione Treansferase". Toxicology Methods, 7:1051 (1997).
What is the the source of the glutathione reductase used as the standard?
The source of standard in the kit is S. cerevisiae.
Can frozen tissues be used in this assay?
The assay protocol is validated for isolating Glutathione Peroxidase from fresh tissue. We have not tested frozen tissue in this assay. Fresh tissue is recommended because single cell suspension from tissue extracts is needed to eventually prepare cytosolic extracts. It is very difficult to make single cell extracts from frozen tissue and most of the cells have likely already lysed from freezing. We recommend isolating the cytosolic extracts prior to freezing the samples.
Which kind of plate can be used for a substitute plate?
A 96-well, flat bottom ELISA plate can be substituted for the plates supplied.
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