Product Introduction
This kit can detect the content of GSH (reduced glutathione) and GSSG (oxidized glutathione disulfide) respectively. Glutathione (glutathione) is a small peptide composed of three amino acid residues. Its full name is glutamyl-cysteinyl-glycine, abbreviated as glutathione. Since the sulfhydryl group (SH) on cysteine is the active group of glutathione, it is often abbreviated as G-SH or GSH. Glutathione includes two forms: reduced glutathione and oxidized glutathione disulfide. Since oxidized glutathione is formed by the dehydrogenation of two GSH groups through sulfhydryl groups, it is often abbreviated as G-S-S-G or GSSG. Reduced glutathione is the main source of sulfhydryl groups in most living cells. It plays an important role in maintaining the appropriate redox state of sulfhydryl groups in proteins and is a key antioxidant in animal cells. Typically 90-95% of total glutathione is reduced glutathione.
Product Features
1.The detection limit of this kit is 0.5μM, and it can detect the content of GSH and GSSG in animal tissues, plasma, red blood cells, cultured cells or other appropriate samples.
2.This kit provides protein removal reagent M, which can more accurately measure samples containing protein.
Principle
GSSG is reduced to GSH by glutathione reductase, and GSH can react with the chromogenic substrate DTNB to produce yellow TNB and GSSG. When the reaction system is properly prepared and the two reactions are combined, the total glutathione (GSSG+GSH) is equivalent to a rate-limiting factor in color production, and the amount of total glutathione determines the amount of yellow TNB formed. Therefore, the amount of total glutathione can be calculated by measuring the absorbance value at 412nm. Use appropriate reagents to first remove GSH from the sample, and then use the above reaction principle to determine the GSSG content. The GSH content can be calculated by deducting the GSSG content from the total glutathione (GSSG+GSH).
The specific reaction principle of this kit is as follows:
The two reactions are combined:
Components
Serial number | Components | Size(100T) | Storage |
Reagent 1 | Total Glutathione Assay Buffer | 60ml | -20℃ |
Reagent 2 | Glutathione reductase | 150μl | -20℃ |
Reagent 3 | Oxidized Glutathione (GSSG) | 5mg | -20℃, GSSG is prepared into solution, packaged, and stored at -20ºC for at least 3 months |
Reagent 4 | DTNB | 4.5mg | -20℃, after DTNB is dissolved in DMSO, it is packaged and stored at -20ºC for at least 3 months. |
Reagent 5 | Protein Removal Reagent M | 1g | -20℃, Protein Removal Reagent M can only be used on the same day after being prepared into solution. |
Reagent 6 | NADPH | 4mg | -20°C, dissolve NADPH, aliquot, and store at -70°C. |
Reagent 7 | DMSO | 1.5ml | -20℃ |
Reagent 8 | GSH Scavenging reagents | 500μl | -20℃, GSH scavenging reagent solution must be freshly prepared for use. |
Consumable 1 | Microplate(96 wells) | 1 plate | RT |
Consumable 2 | Plate Sealer | 2 pieces | RT |
Storage
The unopened kit can be stored at -20℃ for 12 months.
Notes
This product is intended for scientific research use only by professionals and must not be used for clinical diagnosis or treatment, in food or drugs, or stored in ordinary residences.
