Glutathione Peroxidase (GPx) Activity Assay

NWLSS | Northwest Life Science Specialties, LLC.
Glutathione Peroxidase (GPx) Activity Assay
NWK-GPX01 $315.00 each

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A kinetic, rate based assay kit for quantification of glutathione peroxidase (GPx) in a variety of samples including blood products, tissues and cultured cells. This assay will work with any species.

Introduction

Glutathione Peroxidase (GPx) (EC 1.11.1.9) enzymes belong to a family of selenoproteins whose function is to catalyze the reduction of various peroxides. Four types of GPx (GPx-1 to 4) have been identified in mammalian species. GPx-1 is the classic intracellular form while GPx-2 to 4 are gastrointestinal, plasma and phospholipid hydroperoxide metabolizing forms respectively.

Test Principle

This assay is an adaptation of the method of Paglia and Valentine. Glutathione peroxidase (GPX) catalyzes the reduction of an organic peroxide (ROOH), oxidizing reduced glutathione (GSH) to form disulfde glutathione (GSSG). The oxidized glutathione is then reduced by glutathione reductase (GR) and b-nicotinamide adenine dinucleotide phosphate (NADPH) forming NADP+ (resulting in decreased absorbance at 340 nm) and recycling the GSH. Because glutathione peroxidase (GPx) is limiting, the decrease in absorbance at 340 nm is directly proportional to the glutthione (GPx) concentration.

Can Glutathione Peroxidase (GPX) be detected in cell culture supernatant?

Glutathione peroxidase (GPx) activity is detectable in the supernatant from homogenates of cultured cells but is likely not present in sufficient quantities in the cell culture medium to be detectable.

Question: Can I use the same deproteinated sample I am testing for Glutathione (GSH) to test for Glutathione Peroxidase (GPx)?

No. Glutathione peroxidase (GPx) is a protein and will be removed by deproteination. It would be necessary to split the sample prior to deproteination then test GPx in the non-deproteinated sample.
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For Research Use Only

Catalog Number: NWK-GPX01
Format: 192 tests presented as 2 X 96 well microtiter plate assays. Assay can also be performed in cuvettes with measurement via spectrophotometer for a 100 test yield per package
Sample Requirements: Tissues and cell lysates
Specificity: Glutathione Peroxidase
Sensitivity: LLD = 5.0 mU/mL by plate reader LLD = 1.0 mU/mL by spectrophotometer
Intended Use: Quantitative measurement of GPx activity
Storage and Stability: 12 months from date of manufacture if stored at 2-8C
Kit Contents: NADPH Reagent (containing GSH)
NADPH Diluent
H2O2 Reagent
Assay Buffer
96 Well Microplates
Random Citation | NWK-GPX01

Ameliorative effect of zinc oxide and silver nanoparticles on antioxidant system in the brain of diabetic rats
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