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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