Hydrolysis of acetylthiocholine iodide/butyrylthiocholine chloride was monitored by the formation of the yellow 5-thio-2-nitrobenzoate anion, as a result of the reaction of DTNB
with thiocholines, catalyzed by enzymes (at 412 nm), utilizing a 96-well microplate reader (VersaMax Molecular Devices, Sunnyvale, CA, USA).
The amount of GSH in the samples was determined by using DTNB
l3]C NMR, proton and carbon-13 nuclear magnetic resonances; ACTI, acetylthiocholine iodide; MTT, thiazolyl blue tetrazolium bromide; DTNB
, 5,5-dithiobis [2-nitrobenzoic acid]; NaOH, sodium hydroxide; DMSO, dimethyl sulfoxide; EMEM, Eagle's Minimum Essential Medium; FBS, fetal bovine serum; C[O.
Cys, GSH, and DTNB
were purchased from Sigma-Aldrich (Sigma-Aldrich Co.
combines with glutathione to generate a yellow product with maximal absorbance at 412 nm (Cuvette Assay for GSH/GSSG, Oxford Biomedical Research, Oxford, MI).
GSH content in root and shoot of maize was analyzed spectrophotometrically with DTNB
used as chromogenic agent.
Se adicionaron a cada cavidad, 25 [micro]L de la solucion de yoduro de acetilcolina (ATCI), 125 [micro]l de la solucion de DTNB
(5',5-dithio-bis-(2-nitrobenzoate, Sigma), y 50 [micro]L de Tris/HCl (50mM) con albumina serica bovina.
Here the DTNB
(5, 5'-dithiobis- (2- nitrobenzoic acid) react with GSH to form GSH-TNB that absorb at 412 nm.
Finalmente, para la ACH y ACH in, se colocaron 100 [micro]l del HM mas 100 [micro]l de ATCH (mezcla de 75mg de ATCH/100ml de BFP) y 100 [micro]l de DTNB
(mezcla de 13mg de DTNB
/100ml de BFP).
were added in it and absorbance was taken at 412nm.
was used as a chromogen, and it formed a yellow-coloured complex with SH groups.
Disodium edetate (Reidel Dehan AG sleeze Hannover) sodium chloride NaCl (Merck) sodium hydroxide (Fluka AG) chloroform (Merck) DTNB
of Sigma company cadmium acetate (Sigma) Reduced form of GSH of Fluka HCl 35% (Kolchlight) distilled water double refined potassium dihydrogen phosphate (Merck) water for injection (Elexor laboratories).