ELISA/assay

Glutathione Reductase Activity Kit

SKT-204-96

96 well

366.00 USD

Glutathione Reductase Activity Kit

Fluorometric detection of glutathione reductase activity by the amount of GSH generated from the reduction of GSSG

Glutathione Reductase

366.00 USD

USD

Cancer Oxidative Stress

γ-L-Glutamyl-L-cysteinylglycine Activity Kit (2S)-2-Amino-5-[[(2R)-1-(carboxymethylamino)-1-oxo- 3-sulfanylpropan-2-yl]amino]-5-oxopentanoic acid Activity Kit

96 well

Assay Kits

Activity Kits

StressXpress®

Fluorometric Assay

Direct Enzyme Activity Assay

Enzyme Activity

Microplate

Fluorometric assay used to determine glutathione reductase activity by the amount of GSH generated from the reduction of GSSG, in samples.

The Glutathione Reductase Activity kit is designed to quantitatively measure glutathione reductase (GR) activity in a variety of samples. A GR standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. The kit utilizes a proprietary non-fluorescent molecule that will covalently bind to the free thiol group on GSH generated in the reduction of oxidized glutathione (GSSG) to yield a highly fluorescent product. After mixing the sample or standard with StressXpress® Detection Reagent and incubating at room temperature, the fluorescent product is read at 510 nm in a fluorescent plate reader with excitation at 390 nm. Background thiol content is read first after 5 minutes, followed by addition of GSSG and NADPH which uses the standard or sample GR to convert the oxidized glutathione, GSSG, into free GSH, which then reacts with the StressXpress® Detection Reagent to yield the signal related to GR activity The activity of GR in the sample is calculated from the generated signal. We have provided a 96 well plate for measurement but this assay is adaptable for higher density plate formats. The end user should ensure that their HTS black plate is suitable for use with these reagents prior to running samples.

SKC-204A SKC-204B SKC-204C SKC-204D SKC-204E SKC-204F SKC-204G SKC-204H

Black Half Area 96 Well Plate Glutathione Reductase Standard StressXpress® Detection Reagent Dry DMSO Assay Buffer Concentrate NADPH NADPH Diluent Oxidized Glutathione

1 Plate 40 µl 1 vial 2 ml 50 ml 1 vial 5 ml 3 ml

Intra Assay Precision: Five native samples diluted in Assay Buffer and run in replicates of 16 in an assay. The mean and precision of the calculated GR activities were: Sample 1- 3.35 mU/mL, 2.7% CV Sample 2- 2.38 mU/mL, 5.6% CV Sample 3- 1.75 mU/mL, 3.9% CV Sample 4- 0.56 mU/mL, 3.7% CV Sample 5- 0.27 mU/mL, 3.6% CV Inter Assay Precision: Five native samples were diluted in Assay Buffer and run in duplicates in twenty-two assays run over multiple days by four operators. The mean and precision of the calculated GR activities were: Sample 1- 3.35 mU/mL, 5.0% CV Sample 2- 2.36 mU/mL, 12.6% CV Sample 3- 1.64 mU/mL, 6.8% CV Sample 4- 0.62 mU/mL, 10.67% CV Sample 5- 0.27 mU/mL, 10.5% CV

Human

Hu

Serum EDTA Plasma Heparin Plasma Erythrocytes Cell Lysates

41 samples in duplicate

20 Minutes

0.009 mU/ml

0.156 - 5 mU/ml

4ºC

Blue Ice

Glutathione Reductase Activity Kit (StressMarq Biosciences, Canada, Cat # SKT-204-96)

Glutathione reductase (GR) plays an indirect but essential role in the prevention of oxidative damage within the cell by helping to maintain appropriate levels of intracellular glutathione (GSH). GSH, in conjuction with the enzyme glutathione peroxidase (GP), is the acting reductant responsible for minimizing harmful hydrogen peroxide cellular levels (1). The regeneration of GSH is catalyzed by GR (2). GR is an ubiquitous 100-120 kDa dimeric flavoprotein that catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione, using ß-nicotinamide dinucleotide phosphate (NADPH) as the hydrogen donor (3). Molecules such as NADPH act as hydride donors in a variety of enzymatic processes. NADPH has been suggested to also act as an indirectly operating antioxidant, given its role in the re-reduction of GSSG to GSH and thus maintaining the antioxidative power of glutathione.

1. Meister A. (1994) J Biol Chem. 269(13): 9397-400. 2. Massey V., & Williams C. (1965) J Biol Chem. 240(11): 4470-80. 3. Andersen H., Nielsen J., Nielsen F., & Grandjean P. (1997) Clin Chem. 43(4): 562-8.

Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Kit Booklet MSDS

SKT-204_Glutathione_Reductase_Activity_Kit_End-Point_Assay_Standard_Curve_Fig2.png SKT-204_Glutathione_Reductase_Activity_Kit_Chemical_Equation_of_GR_Reaction_Fig1.png SKT-204_Glutathione_Reductase_Activity_Kit_Linearity_Recovery_Graph_Fig3.png
SKT-204_Glutathione_Reductase_Activity_
Kit_Continuous_Kinetic_Assay_Standard_C
urve_Fig4.png

Graph of the Continuous Kinetic Assay Typical Standard Curve for the Glutathione Reductase Activity Kit StressXpress® - SKT-204 Graph of the End-Point Assay Typical Standard Curve for the Glutathione Reductase Activity Kit StressXpress® - SKT-204 Chemical Equation of the Glutathione Reductase Reaction for the Glutathione Reductase Activity Kit StressXpress® - SKT-204 Graph of the Linearity Recovery for the Glutathione Reductase Activity Kit StressXpress® - SKT-204