|Product name||CheKine™ Glutathione S-Transferase (GST) Colorimetric Assay Kit|
|Applications notes||GST catalyzes the binding of GSH to CDNB. The conjugation is accompanied by an increase in absorbance at 340 nm. The rate of increase is directly proportional to the GST activity in the sample.|
|Kit components||• Assay Buffer
|Features & Benefits||• Determination of GST activity in serum, plasma, tissue/cell lysates and other biological fluids.
• A broad range linearity: 2-76 U/L.
• Sensitivity down to 2 U/L.
|Usage notes||• 96 Well Clear Flat Bottom UV-Transparent Microplate is needed.
• The sample preparation should be carried out on ice.
• If the experiment is not carried out immediately, the samples can be kept at -80oC for 1 month.
• In the detection of GST activity in cells, the cell number must be between 3-5 x 106, and the extraction of GST in cells can be followed by Assay Buffer grinding or ultrasonic treatment, cells cannot be treated by lysate.
• Pre-test is needed before experiment, two or three samples with large difference should be diluted into different concentrations to test.
• If the results collected within 5 minutes are not linear, or the sample absorbance value is greater than 1, it is suggested that the sample should be diluted with distilled water and the result should be multiplied by the dilution factor.
• The reaction temperature has influence on the result. Keep the temperature at 25°C (for general species) or 37°C (for mammals).
|Storage instructions||Storage at -20°C and Keep from light immediately upon receipt. Kit has a storage time of 12 months from receipt. Refer to list of materials supplied for storage conditions of individual components.|
|Shipping||Gel pack with blue ice.|
|Precautions||The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.|
|Background||Glutathione S-transferases (GSTs) are ubiquitous multifunctional enzymes, which play a key role in cellular detoxification. The enzymes protect cells against toxicants by conjugating them to glutathione, thereby neutralizing their electrophilic sites, and rendering the products more water-soluble. The glutathione conjugatesare metabolized further to mercapturic acid and then excreted. GST has the activity of GSH-Px, also called non-Se GSH-Px, which has the function of repairing oxidative damage of macromolecule such as DNA, protein and so on. GST catalyzed reaction decreased GSH content, but did not increase GSSG content. Based on their sequence homology, substrate specificity and immunological cross-reactivity, GSTs have been grouped into species-independent classes of isozymes. These classes are comprised of both cytosolic and microsomal enzymes.|
Fig. GST kinetic curve. The y-axis is the absorbance and the x-axis is the time (min).