Wednesday, August 26, 2015

Procedure for glutathione estimation

Glutathione is tripeptide, L-γ-glutamyl-L-cysteinylglycine present in high concentrations in most cell types. It is most powerful and naturally occurring antioxidant making it critically important in eliminating detoxification and free radicals. In breast cancer, the GSH concentration is twice more compared to normal patients.

1.810 ml of distilled water is added to 0.2 ml of the haemolysate.  0.01 ml of lysate is added to 3 ml of ferricyanide-cyanide reagent for hemoglobin estimation, and 3 ml of precipitating solution is added to the remaining haemolysate. After standing for 10 minutes, the mixture is filtered through a medium or coarse grade of filter paper. One milliliter of filtrate is added to 4 ml of 0.3 M Na2 HPO4 solutions in a cuvette with a critical volume of 10 ml or less. It is read at 412 nm against a blank prepared by adding one ml of distilled water to 4 ml of 0.3 M Na2 HPO4. Optical density reading is taken after 0.5 ml of DTNB is added to the blank cuvette and the cuvette containing filtrate

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