Glutathione (GSH) Fluorescent Detection Kit

Research Areas

Documentation

Specifications

• Assay Type Detection Kit
• Sample Types Whole Blood, Serum, Plasma (EDTA and Heparin), Erythrocytes, Urine, Cell Lysates, Tissue
• Sensitivity 45 nM Free GSH, 48 nM Total GSH
• Species GSH is identical across species
• Assay Duration 30 Minutes
• Samples/Plate 39 in Duplicate
• Readout Fluorescent, 510 nm emission / 370-410 nm excitation
• Standard Curve
• Description

  Assay Principle: 

  The Glutathione (GSH) Fluorescent Detection Kit quantitatively measures GSH levels in whole blood, serum, plasma (EDTA and Heparin), erythrocytes, urine, cell lysates, and tissue. The Glutathione (GSH) Fluorescent Detection Kit is a Detection Kit with a run time of 30 minutes. Please read the complete kit insert for more information before performing this assay.

  Use our provided Glutathione standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a black microtiter plate. Add ThioStar® Detection Reagent to each well tapping the plate to ensure sufficient mixing of reagents. Then incubate the mixture covered at room temperature for 15 minutes. The fluorescent reaction occurs between the ThioStar® Detection Reagent and the GSH within the sample or standard. 

  After the 15-minute incubation, use a plate reader to detect and record the generated fluorescent signal. Use the intensity and the standard curve to calculate the GSH concentration in the samples.

  Background:

  Glutathione (L-γ-glutamyl-L-cysteinyl glycine; GSH) is the highest concentration of non-protein thiol in mammalian cells and is present in concentrations of 0.5 – 10 mM. GSH is a tripeptide molecule. It contains a peptide linkage between the amine group of cysteine and the carboxyl group of the glutamate side chain. It is an antioxidant, preventing damage to important cellular components caused by reactive oxygen species. Enzymes convert Glutathione to its oxidized form, glutathione disulfide (GSSG). By serving as an electron donor, Glutathione reduces disulfide bonds formed within cytoplasmic proteins to cysteines. Its reduced form is most common since glutathione reductase is constitutive and inducible upon oxidative stress. Investigators use the ratio of reduced Glutathione to oxidized Glutathione within cells to measure cellular toxicity.

• Structure