Reduced glutathione

Streptozotocin was procured from Hi-media (Mumbai, India). Ammonium sulphate, 1-chloro-2,4-dinitrobenzene, ethylenediaminetetraacetic acid, 2,4-dinitro-phenyl-hydrazine, 5,5-di-thio-bi(2-nitrobenzoic acid), potassium dihydrogen phosphate, N-ethylmaleimide, reduced nicotinamide adenine dinucleotide, nitro blue tetrazolium, sodium pyrophosphate, phenazinemethosulphate, thiobarbituric acid, reduced glutathione, sodium azide, trichloro acetic acid and 5-thio-2-nitrobenzoic acid were obtained from Sisco Research Laboratory (India). Bradford reagent, antibodies and bovine serum albumin were procured from Sigma-Aldrich (St. Louis, USA). The kits for different assays for different biochemical parameters were purchased from Span diagnostic Ltd., India. All other reagents and chemicals used were of analytical grade.

The technical grade (99.6% pure) carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl methylcarbamate) in powder form was a kind gift of Rallis India Limited, Bangalore, India. Edible groundnut oil was purchased from the local market. Pyrogallol, reduced glutathione (GSH), 1-chloro-2,4-dinitrobenzene (CDNB), acetylthiocholine-iodide (ATI), 3,5-dithionitrobenzoic acid (DTNB), and bovine serum albumin (BSA) were purchased from Sisco Research Laboratories Pvt. Ltd., Mumbai, India. Curcumin was purchased from Sigma-Aldrich Inc., USA. All other chemicals used in the study were of analytical grade.

The 1-chloro-2,4-dinitrobenzene, ammonium sulphate, 2,4-dinitrophenylhydrazine, ethylene diaminetetraacetic acid, 5,5-dithiobis(2-nitrobenzoic acid), N-ethylmaleimide, nitroblue tetrazolium, reduced nicotinamide adenine dinucleotide, potassium dihydrogen phosphate, phenazinemethosulphate, sodium pyrophosphate, reduced glutathione, sodium azide, thiobarbituric acid, 5-thio-2-nitrobenzoic acid and trichloro acetic acid were purchased from Sisco Research Laboratory, Mumbai, India. Bradford reagent, bovine serum albumin and cadmium chloride were procured from Sigma-Aldrich Chemical Company, St. Louis, USA. All antibodies for immunoblotting were purchased from Sigma-Aldrich Chemical Company St. Louis, USA. HPLC grade solvents (acetic acid, formic acid, methanol and acetonitrile) were procured from Merck, Mumbai, India.

The chemicals utilized during study: ammonium sulphate, 2,4-dinitro-phenyl-hydrazine, 1-chloro-2,4-dinitrobenzene, ethylenediaminetetraacetic acid, 5,5-di-thio-bi(2-nitrobenzoic acid), N-ethylmaleimide, nitro blue tetrazolium, reduced nicotinamide adenine dinucleotide, potassium dihydrogen phosphate, phenazinemethosulphate, sodium pyrophosphate, reduced glutathione, sodium azide, thiobarbituric acid, 5-thio-2-nitrobenzoic acid and trichloro acetic acid were procured from Sisco Research Laboratory, India. Bovine serum albumin, Bradford reagent and Pb-acetate were purchased from Sigma-Aldrich, St. Louis, USA. All primary antibodies (produced in rabbit) viz. anti-Bcl-2 (dilution 1:1000), anti-Bad (dilution 1:3000), anti-Cyt C (dilution 1:1000), anti-Apaf-1 (dilution 1:1000), anti-caspase 3 (dilution 1:1000), anti-caspase 8 (dilution 1:1000), anti-caspase 9 (dilution 1:1000), anti-Fas (dilution 1:2000), anti-Bid (dilution 1:1000) and anti-actin (dilution 1:3000) for immunoblotting were purchased from Sigma-Aldrich Chemical Company St. Louis, USA. Appropriate HRP conjugated secondary antibody (dilution 1:3000) produced in goat was also purchased from Sigma-Aldrich Chemical Company St. Louis, USA. Methanol, formic acid, acetic acid and acetonitrile were obtained from Merck, India.