Unit Price (USD): 1 kit $160.55

Bulk Price/unit (when you buy 5 or more): $128.44

Description:
Provides reagents and methods to simply and quickly measure glutathione transferase activity in live cells, tissues or cell lysate samples

Application:
Glutathione (GSH), is a tripeptide (g-glutamyl-cysteinyl-glycine) that represents the major free thiol in most living cells. It is involved in many biological processes including detoxification of xenobiotics, removal of hydroperoxides, and maintenance of the oxidation state of protein sulfhydryls. It is the key antioxidant present in animal tissues, and diminished glutathione levels have been observed in the early stages of apoptosis. Older methods for determining glutathione utilize 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB or Ellman's reagent) in an oxidation or oxidation-reduction scheme. But these methods have been largely replaced by a widely used method for determining GSH in living cells. The Live Cell Glutathione Transferase Assay Kit allows intracellular glutathione S-transferase detection by simply adding a fluorogenic reagent mCB to the cell culture medium or lysate to form GSH-mCB complexes. The levels of GSH can be measured fluorometrically. Unlike other bimanes such as monobromobimane, monochlorobimane appears to form an adduct exclusively with GSH. This membrane-permeant reagent has been used to measure GSH in live cells and in flow cytometry systems. This procedure has been used to measure GSH content of cultured neural cells and in tissue homogenates and, indeed, several laboratories have used this approach to measure the GSH content of the cytosolic fraction of liver or in intact tissues. It has been found that monochlorobimane readily enters cells to form a fluorescent GSH mono-chlorobimane adduct that can be measured fluorometrically and that this reaction is catalyzed by glutathione S-transferase. The absorption and emission maxima of the complex formed is 380nm and 461nm, respectively. The kit contains enough reagents for up to 100 labelings, including lysis buffer, solvents, and a detailed protocol for use. Please contact our technical services department for further information.

References:

1. Fernandez-Checa, J. C., Kaplowitz, N. (1990) "The use of monochlorobimane to determine hepatic GSH levels and synthesis." Anal. Biochem. 190: 212-219.
2. H. Kamencic, A. Lyon, P. G. Paterson, B. H. J. Juurlink (2000) "Monochlorobimane Fluorometric Method to Measure Tissue Glutathione" Analytical Biochemistry 286: 35-37.
3. Reichelt, W., Stabel-Burow, J., Pannicke, T., Weichert, H., Heinemann, U. (1997) "The glutathione level of retinal Müller glial cells is dependent on the high-affinity sodium-dependent uptake of glutamate." Neuroscience 77: 1213-1224.
4. Devesa, A., Oconnor, J. E., Garcia, C., Puertes, I. R., Vina, J. R. (1993) "Glutathione metabolism in primary astrocyte cultures: flow cytometric evidence of heterogeneous distribution of GSH content." Brain Res. 618: 181-189.
5. Butcher EC, Scollay RG, Weissman IL., J Immunol Methods 37(2), (1980) 109-121.
6. Kannan, R., Tang, D., Mackic, J. B., Zlokovic, B. V., Fernandez-Checa, J. C. (1993) "A simple technique to determine glutathione (GSH) levels and synthesis in ocular tissues as GSH-bimane adduct: application to normal and galactosemic guinea-pigs." Exp. Eye Res. 56: 45-50.