Ethylenediaminetetraacetic acid

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.

Agarose, 1-chloro-2,4-dinitrobenzene, 2,2-diphenyl-1-picrylhydrazyl (DPPH), N-ethylmaleimide, glutathione reductase (GR), L-leucine p-nitroanilide, metaphosphoric acid, NaNO₂, Roswell Park Memorial Institute 1640 (RPMI) medium, Triton X-100, 2,4,6-tris(2-pyridyl)-s-triazine, p-nitrophenyl phosphate, γ-glutamyl p-nitroanilide, o-phthalaldehyde and NaNO₂ were from Sigma Aldrich, USA. Bovine serum albumin, diphenylamine, 5,5'-dithiobisnitrobenzoic acid (DTNB), ethylenediaminetetraacetic acid (EDTA), glucose 6-phosphate, proteinase K, reduced (GSH) and oxidized (GSSG) glutathione, hydrogen peroxide (H₂O₂), reduced and oxidized nicotinamide adenine dinucleotide phosphate (NADPH and NADP⁺), sodium dodecyl sulfate (SDS), sorbitol, tris(hydroxymethyl)aminomethane (Tris), pyrogallol, 2,4-dinitrophenylhydrazine and xylenol orange were from Sisco Research Laboratories, India. Thiobarbituric acid and trichloroacetic acid were from Himedia Laboratories (Mumbai, India).
All other chemicals used were of analytical grade.

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.

Bovine serum albumin (BSA), Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), Bradford reagent, Collagenase type I, antibodies for immunoblotting were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO), USA. Solvents (HPLC grade) were obtained from Merck, Mumbai, India. Kits for measurement of serum biochemical parameters were purchased from Span diagnostic Ltd., India. 1-Chloro-2,4-dinitrobenzene, ammonium sulphate, sodium arsenite (NaAsO₂), 2,4-dinitrophenylhydrazine, ethylene diaminetetraacetic acid, 5,5-dithiobis(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 purchased from Sisco Research Laboratory, Mumbai, India.

Bovine serum albumin (BSA), glycine, protease inhibitor cocktail, RA and Tween 20 were from Sigma-Aldrich (St. Louis, MO, USA). CP was procured from Dr Reddy’s Laboratories (Hyderabad, India). Primary and secondary antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and Elabscience (Houston, TX, USA). Eosin, hematoxylin, Ponceau S, β-mercaptoethanol and Triton X-100 were purchased from Hi-Media Laboratories Pvt. Ltd. (Mumbai, India). Acrylamide, bisAcrylamide, ethylene-diaminetetraacetic acid and tris-hydroxymethylaminomethane were purchased from Sisco Research Laboratories Pvt. Ltd. (Mumbai, India). Horseradish peroxidase was from E. Merck Ltd. (Mumbai, India). Other routine chemicals and reagents were purchased from either Sigma-Aldrich or accredited local vendors.