GSH and GSSG Assay Kit
$211.00
All products have special prices for bulk purchase, please contact us for more details if required.
Cat. No.: GSHG-100 (for 100T)
Description
GSH and GSSG Assay Kit is a simple and easy-to-use assay kit that allows for the separate detection of GSH (reduced glutathione) and GSSG (oxidized glutathione disulfide) levels.
Glutathione (GSH) is a small peptide composed of three amino acid residues. Its full name is glutamyl-cysteinyl-glycine, and its abbreviation is glutathione. The sulfhydryl group (SH) on the cysteine residue is the active group of glutathione, hence it is commonly referred to as G-SH or GSH. Glutathione exists in two forms: reduced glutathione (GSH) and oxidized glutathione disulfide (GSSG). GSSG is formed by the oxidation of two GSH molecules through disulfide bond formation, and it is often represented as G-S-S-G or GSSG. GSH is the primary source of thiols in most active cells and plays a crucial role in maintaining the appropriate redox state of proteins. It is also a key antioxidant in animal cells. Typically, 90-95% of total glutathione is in the reduced form (GSH).
In the assay, glutathione reductase is used to convert GSSG to GSH. GSH then reacts with the colorimetric substrate DTNB to produce a yellow-colored product, TNB, and GSSG. By appropriately setting up the reaction system and combining the two reactions, the total glutathione (GSSG+GSH) becomes the limiting factor for color production, and the amount of total glutathione determines the formation of yellow TNB. Therefore, the amount of total glutathione can be calculated by measuring the absorbance at 412 nm (A412). By depleting GSH in the sample using an appropriate reagent and applying the above reaction principle, the content of GSSG can be determined. Subtracting the GSSG content from the total glutathione (GSSG+GSH) allows for the calculation of GSH content.
The GSH and GSSG Assay Kit can detect the levels of GSH and GSSG in animal tissues, plasma, red blood cells, cultured cells, or other appropriate samples.
The kit provides a protein removal reagent (M) to accurately measure the amounts of GSH and GSSG in protein-containing samples.
The detection limit of the assay kit is 0.5 μM. Each kit allows for 100 tests, enabling the measurement of total glutathione or GSSG content in 100 samples, or the determination of GSH and GSSG content in 50 samples each.
Additional Notes
- The assay involves redox reactions, and all oxidizing agents or reducing agents can interfere with the assay. In particular, DTT, mercaptoethanol, and other thiol-containing reagents can significantly interfere with the assay and should be avoided as much as possible.
- It is important to strictly control the temperature and reaction time during the assay. Otherwise, a standard curve must be prepared for each assay.
- Reagents such as NADPH are not very stable, so it is crucial to follow the instructions carefully to prevent inactivation.
- The protein removal reagent (M) solution must be freshly prepared and used on the same day. The GSH clearing reagent should also be freshly diluted before use.
- The protein removal reagent (M) may be difficult to dissolve, but vigorous vortexing and mild heating (not exceeding 37°C) can help facilitate dissolution.
- DMSO may solidify at lower temperatures such as 4°C or during ice baths. It can be thawed by placing it in a water bath at 20-25°C until fully dissolved before use.
- This product is intended for scientific research purposes by professionals only. It should not be used for clinical diagnosis or treatment, or for food or drug purposes. It should not be stored in residential areas.
- For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling the product.
Storage
Store at -20°C, valid for one year. After GSSG is prepared as a solution, it should be properly aliquoted and stored at -20°C for at least 3 months. DTNB dissolved in DMSO should also be properly aliquoted and stored at -20°C for at least 3 months. After preparing the protein removal reagent (M), it should be used on the same day. NADPH, once dissolved, should be properly aliquoted and stored at -70°C. The GSH Clearing Assay Buffer and GSH Clearing Reagent, when diluted, should be freshly prepared and used.
Only for research and not intended for treatment of humans or animals
Journals Using SBS Genetech Products Universities Using SBS Genetech Products
SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory
