Glutathione Reductase Assay Kit with DTNB
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Cat. No.: GRAKD-100 (for 100T)
Glutathione Reductase Assay Kit with DTNB is a simple and convenient assay kit used to detect the activity of Glutathione reductase (GR) in cells, tissues, or other samples using a colorimetric method. Glutathione reductase is distributed in many tissues and helps maintain adequate levels of reduced glutathione (GSH) within cells. GSH can eliminate free radicals and some organic peroxides, or serve as a substrate for Glutathione peroxidase to eliminate certain peroxides.
Glutathione reductase can reduce oxidized glutathione (GSSG) to generate reduced glutathione (GSH). GSH can react with the colorimetric substrate DTNB to produce a yellow product called TNB along with GSSG. The amount of TNB generated can be measured by monitoring the absorbance at 412 nm (A412). By setting up the appropriate reaction system, combining the two reactions mentioned above, and when there is an excess of GSSG/GSH relative to GR, glutathione reductase becomes the rate-limiting factor in the reaction system. In this case, the amount of TNB formed and the activity of glutathione reductase are linearly correlated. Therefore, by measuring A412, the activity level of glutathione reductase can be calculated.
One kit allows for 100 tests.
- The assay kit involves oxidation-reduction reactions, therefore, all oxidizing or reducing agents can interfere with the assay. If reducing agents in the sample cannot be avoided, such as DTT, mercaptoethanol, etc., the total concentration of these agents should be kept below 0.1 mM. DTT at a concentration of 0.15 mM can inhibit 40% of the enzyme activity. Additionally, sodium sulfate, ammonium sulfate, and ferricyanide can also interfere with the assay. Please avoid their presence as much as possible.
- Samples can be tested immediately or stored at -70°C for future analysis.
- It is crucial to strictly control the temperature at 25°C during the reaction to minimize errors.
- NADPH is not very stable, so it is important to follow the instructions carefully to prevent its inactivation.
- This product is intended for use by professionals in scientific research only. It should not be used for clinical diagnosis or treatment, or for food or drug purposes. It should not be stored in a regular household setting.
- For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling the kit.
Store at -20°C, valid for one year. After dissolving NADPH, it is recommended to aliquot and store at -70°C. It can be stored at 4°C for one day, but after one week of storage at -20°C, NADPH may degrade by more than 10%. GSSG and DTNB, after preparation as solutions, should be aliquoted appropriately and stored at -20°C.
Only for research and not intended for treatment of humans or animals
SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory