The DetectX® Glutathione (GSH) Fluorescent Detection Kit utilizes a proprietary non-fluorescent molecule, ThioStar®, that covalently binds to the free thiol group on GSH to yield a highly fluorescent product.

检测类型: Detection Kit
样本类型: Serum, Plasma (EDTA and Heparin), Cell Lysates, WB, RBCs, Urine, Tissue
灵敏度: 45 nM (Free GSH), 48 nM (Total GSH)
适用物种: GSH在不同物种间是相同的
检测时间: 15分钟
样品数/板: 183 in Duplicate (Free GSH), 185 in Duplicate (Total GSH)
读数: Fluorescent, 510 nm emission / 370-410 nm excitation

检测原理:
The Glutathione (GSH) Fluorescent Detection Kit (384-Well Plate) quantitatively measures GSH levels in whole blood, serum, plasma (EDTA and Heparin), erythrocytes, urine, cell lysates, and tissue. The Glutathione (GSH) Fluorescent Detection Kit (384-Well Plate) 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.

背景:
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.