Dithiobis 2 nitrobenzoic acid

4,2-Hydroxyethyl, 1-piperazineethanesulfonic acid (HEPES), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carbo 2-carboxylic acid (Trolox), 3-(N-morpholino) propane sulfonic acid (MOPS), dimethyl sulfoxide (DMSO), D-mannitol, fatty acid-free bovine serum albumin (BSA) fraction V, Thiobarbituric acid (TBA), 3-[4 (link), 5 (link) dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT), Dithiobis-2-nitrobenzoic acid (DTNB), Glutathione (GSH), 2’, 7’-Dichlorofluorescein diacetate (DCFH-DA), Taurine, Malondialdehyde (MDA), Sucrose, KCl, Na₂HPO₄, MgCl₂, Rhodamine 123 (Rh-123), Coomassie brilliant blue, Ethylene glycol-bis (2-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA), Sodium succinate, and Ethylenediaminetetraacetic acid (EDTA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Trichloroacetic acids (TCA), ammonium chloride, and hydroxymethyl amino methane hydrochloride (Tris-HCl) were purchased from Merck (Dardamstd, Germany). All salts for preparing buffer solutions were of analytical grade and prepared from Merck (Dardamstd, Germany).

Fatty acid-free bovine serum albumin (BSA) fraction V, dithiobis-2-nitrobenzoic acid (DTNB), 6-hydroxy-2,5,7,8-tetramethyl chroman-2-carboxylic acid (Trolox), 4,2hydroxyethyl,1-piperazine ethane sulfonic acid (HEPES), thiobarbituric acid (TBA), 2′,7′dichlorofluorescein diacetate (DCFH-DA), taurine (TAU), malondialdehyde (MDA), glutathione (GSH), sodium phosphate dibasic (Na₂HPO₄), sucrose, potassium chloride (KCl), Coomassie brilliant blue, dithiothreitol, ethylene glycol-bis (2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), and ethylenediaminetetraacetic acid (EDTA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Hydroxymethyl aminomethane hydrochloride (Tris-HCl), and trichloroacetic acid (TCA) were obtained from Merck (Darmstadt, Germany). All salts (analytical grade) for preparing buffer solutions were obtained from Merck (Darmstadt, Germany).

Glycine (2-aminoacetic acid), malondialdehyde, thiobarbitu-ric acid, dithiobis-2-nitrobenzoic acid, GSH, 2′,7′-dichloro-fluorescein diacetate (DCFH-DA), and EDTA were provided by Sigma-Aldrich (St. Louis, MO, USA). Trichloroacetic acid (TCA), dithiothreitol (DTT), ferric chloride hexahydrate, 2, 4, 6-Tris (2-pyridyl)-s-triazine (TPTZ), and Tris (hydroxy-methyl) aminomethane hydrochloride were provided by Merck (Darmstadt, Germany).

Ellman method²⁰ (link), modified by adding an inhibitor of BChE activity (tetra-iso-propylpyrophosphoramide, 0.1 mmol/L) or an inhibitor of AChE activity (BW284c51, 20 μmol/L), was employed to determine the activities of AChE and butyrylcholinesterase (BChE). Samples were added to the reaction mixture and make sure the final concentrations of acetylthiocholine iodide at 0.625 mmol/L (ATCh; Sigma) and 5,5-dithiobis-2-nitrobenzoic acid (Sigma) 0.5 mmol/L in 80 mmol/L Na₂HPO₄. Then, the absorbance was measured at 412 nm. Specific enzyme activity was expressed as absorbance units/min/g of protein.