Glutathione reductase gr also known as glutathionedisulfide reductase gsr is an enzyme that in humans is encoded by the gsr gene. Kit measures glutathione peroxidase activity through a coupled reaction with glutathione reductaseeither measuring substrate consumption or overall production, the enzyme assays are used to monitor enzymatic activity. Fr 19 for research use only introduction glutathione reductase gr, ec 1. The assay is based on the glutathione recycling system by dtnb and glutathione reductase fig. Fr 17 for research use only introduction the analyte cellular glutathione peroxidase cgpx, ec 1. Glutathione peroxidase assay kit colorimetric discover more at. Assay designs glutathione peroxidase activity kit is a complete kit for measuring the activity of glutathionedependent peroxidases in cells, tissues, erythrocytes and plasma. Lglutamyllcysteinylglycine is a tripeptide present in the body, and it is involved in antioxidation, drug metabolism, and other as enzyme substrate of glutathione peroxidase, glutathione stransferase, and thiol transferase, etc. Glutathione peroxidase assay kit colorimetric ab102530, gpx reduces cumene. In fact, it is one of the most important antioxidant enzymes in humans. Oxidized glutathione gssg, produced upon reduction of an. In bioassay systems peroxidase assay kit d2pd100, the color 570 nm or fluorescence intensity 530585 nm is a direct measure of the peroxidase activity. Glutathione peroxidase, cellular assay kit cas find msds or sds, a coa, data sheets and more information. Bioassay systems develops and markets innovative and highthroughput assay solutions to satisfy the ever increasing demands of the life sciences industry.
With the exception of phospholipidhydroperoxide gpx, a monomer, all of the gpx. Glutathione colorimetric assay kit bn00529 assay genie. Glutathione reductase gr then reduces the gssg to produce gsh, and in the same reaction consumes nadph. Glutathione peroxidase activity assay with colorimetric method and. It is based on the oxidation of glutathione gsh to oxidized glutathione gssg catalyzed by gpx, which is then. The cayman chemical glutathione peroxidase assay kit measures gpx activity indirectly by a coupled reaction with glutathione reductase gr. Peroxidase assay colorimetric assay for the quantitative determination of glutathione peroxidase gpx activity in tissue homogenates, cell lysates, plasma and erythrocyte lysates. Glutathione peroxidase is the catalyst for glutathione gsh converting hydrogen peroxide h2o2.
The total glutathione peroxidase assay kit is for research purposes only. Glutathione peroxidase an overview sciencedirect topics. To determine glutathione peroxidase reliably, some factors of potential pitfall have to be considered, for example, enzymatic side reactions of substrates especially when crude tissue samples are assayed, high and variable spontaneous reaction rates of substrates, and the peculiar kinetics of the enzyme itself. Monitor the a 340nm until constant, using a suitably thermostatted spectrophotometer. The main differences between the assay procedure described and those. Glutathione is usually present as reduced form gsh, but gsh is converted into its oxidized form gssg by stimulation such as oxidative stress. Gpx oxidizes reduced glutathione in the presence of. This research has been done to investigate a simple, rapid and inexpensive method for. To determine glutathione peroxidase reliably, some factors of potential pitfall have to. Glutathione peroxidase assay kit directly measures nadph consumption in the enzyme coupled reactions. Agecorrelated modifications of copperzinc superoxide dismutase and glutathionerelated enzyme activities in human erythrocytes.
What is glutathione peroxidase ransel assay used for. Enzychromtm glutathione peroxidase assay kit egpx100 quantitative colorimetric glutathione peroxidase determination description glutathione peroxidase transfer 10 gpx, ec 1. Dissolve a 100 unit vial of glutathione peroxidase, catalog number g67, in 1 ml of glutathione peroxidase assay buffer. Collect urine, plasma or saliva and immediately add fou r volumes of 5% wv deproteination reagent. General considerations detection of gsh using the gshglo glutathione assay is the result of combining two chemical reactions.
Glutathione peroxidase gpx glutathione peroxidase gpx is an antioxidant enzyme family. Glutathione peroxidase assay colorimetric assay for the quantitative determination of glutathione peroxidase gpx activity in tissue homogenates, cell lysates, plasma and erythrocyte lysates. Peroxidase assay procedure lin this procedure the ability of the peroxidasegsh system to prevent the oxidative break down of hemoglobin is utilized as a measure of peroxidase activ ity. Prepare enough reaction mixture according to the number of tests to be performed. Glutathione peroxidase activity assay the gpx assay is an indirect, coupled assay for glutathione peroxidase. The enzyme uses glutathione as the ultimate electron donor to regenerate the reduced form of the selenocysteine. It converts reduced glutathione gsh to oxidized glutathione gssg, to reduce lipid hydroperoxides to their corresponding alcohols, or reduce free hydrogen peroxide to water. Gpx functions in the scavenging and inactivating of hydrogen and lipid peroxides, thereby protecting the body against oxidative stress. The gpx assay is an indirect, coupled assay for glutathione peroxidase. The assay takes advantage of concomitant oxidation of nadph. Glutathione reductase is essential for the glutathione redox cycle that. Glutathione peroxidase is a tetramer of four identical subunits, with a molecular weight of 84,000 daltons. The assay kit can determine enzyme activity from different biological sources.
At normal concentrations, selenium has a protective effect against several disease states however this protection is lost at lower concentrations and selenium can be toxic at high concentrations. Identification of the catalytic site of rat liver glutathione peroxidase as selenocysteine. Colorimetric assay for measuring gpx activity in plasma, erythrocyte lysates, tissue homogenates, cell lysate etc. The assay is expected to be most useful for measurement of cellular gpx gpx1. Colorimetric assay for cellular glutathione peroxidase product no. Assay genies glutathione colorimetric assay kit provides a convenient, colorimetric method for analyzing either total glutathione or the reduced form glutathione alone using a microtiter plate reader. Glutathione peroxidase gshpx activity was examined using the procedure of flohe and gunzler 1984. Glutathione peroxidase activity assay gpx assay kit nwlss. Assay for quantitative determination of glutathione and. Glutathione peroxidase protocol the mouse metabolic.
It requires selenium as a cofactor and contains a sele. In the assay, gpx reduce cumene hydroperoxide, and oxidize gsh to gssg. Glutathione peroxidase activity colorimetric assay kit. The glutathione peroxidase positive control is used to assure the reaction is working. Cell biolabs oxiselect glutathione reductase assay kit is a quantitative assay for measuring glutathione reductase activity within plasma, erythrocytes, tissues, and cell lysates. Cellular glutathione peroxidase gpx is a member of a family of. The enzyme glutathione peroxidase gpx was assayed according to the method of flohe and gunzler 1984, with some modifications. In the glutathione peroxidase assay protocol, glutathione peroxidase gpx oxidizes gsh to produce gssg as part of the reaction in which it reduces cumene. Spectrophotometric assay for glutathione reductase product no. Modification for red blood cells as even traces of methemoglobin will falsify the determination of glutathione peroxidase in hemolysate, the conversion of hemoglobin into 12 formation or removal of oxygen radicals 120 comparison of assay procedures for glutathione peroxldase definition of unit gsh0 ram buffer, ph temperature procedure. Colorimetric assay kit used to test for glutathione peroxidase gpx activity in multiple species and sample types. Glutathione peroxidase gpx catalyzes the reduction of hydroperoxides, including hydrogen peroxide, by reduced glutathione and functions to protect the cell from oxidative damage. The generated gssg is reduced to gsh with consumption of nadph by gr.
Provided with the required supplies and reagents for any preferred method, enzyme kinetics and inhibition studies can be successfully completed. In the glutathione peroxidase assay protocol, glutathione peroxidase gpx oxidizes gsh to produce gssg as part of the reaction in which it reduces cumene hydroperoxide. Biovisions glutathione peroxidase assay kit measures glutathione peroxidase gpx activity through a coupled reaction with glutathione reductase gr. The kit employs a simple enzymatic recycling reaction for glutathione. The oxitek total glutathione peroxidase assay kit provides a method of quantifying the activity of total glutathione peroxidase. Glutathione peroxidase cellular activity assay kit cgp1. The purification and properties of glutathione peroxidase. Peroxidase activity is calculated from the od values of the sample, sample blank, calibrator and h 2o wells. Centrifuge the suspension at 12,000 14,000 x g for 10. In biovisions glutathione peroxidase activity assay, gpx reduces cumene hydroperoxide while oxidizing gsh to gssg. The decrease of nadph easily measured at 340 nm is. Sod, cat, and gshpx activities are expressed as units per mg of protein.
To perform the assay the instructions for use provided with the kit have to be used. Peroxidase assay kit measures gpx activity indirectly by a coupled reaction with glutathione reductase gr. General sample preparation protocol a tissue homogenate. Selenium is an essential trace element, involved in the aetiology of a number of diseases. The nwlss glutathione peroxidase assay has been designed to quantify the enzyme activity of glutathione peroxidase. Homogenize the sample in 48 volumes per weight tissue of cold buffer e. This assay takes advantage of glutathione disulfide gssg formed by the enzymatic action of gpx and is regenerated by excess glutathione reductase gr in the assay. Assay sample procedure preparation and detection 10 12. Glutathione peroxidase cellular activity assay kit.
516 1272 1458 992 506 1242 964 1431 1380 1136 226 85 1046 1038 672 579 27 512 1369 313 1143 544 1495 947 697 661 1245 809 1494 661