GST activity was determined against the substrate
CDNB by monitoring thioether formation at 340 nm.
By that point, Precision started shifting traffic to
CDNb, and nearly stopped all traffic to CDNa starting at 12:30 a.m.
[[epsilon].sub.mM] = the extinction coefficient for
CDNB conjugate at
The activities of GST were calculated as described by Habig and Jakoby, (1981) using 1-chloro-2,4-dinitrobenzene (
CDNB) as the substrate.
The reagents [alpha]-naphthyl acetic acid ([alpha]-NA), eserine, fast blue B salt, sodium dodecyl sulfate (SDS), disodium salt of ethylenediamine-tetraacetic acid (EDTA-[Na.sub.2]), dithiothreitol (DTT), reduced glutathione (GSH), nitrobenzene (
CDNB), riboflavin (vitamin B2), N-phenylthiourea, polyvinyl pyrrolidone (PVP), nitro blue tetrazolium (NBT), and methionine (Met) were purchased from Sigma Corporation (Shanghai, China).
Special chemicals like 5' 5' dithio-bis-(2-nitrobenzoic acid) (DTNB), 1- chloro-2, 4-dinitrobenzene (
CDNB), reduced glutathione (GSH), p-phenylenediamine (PPD), were obtained from Sigma chemical co.
Los reactivos utilizados fueron: tampon fosfato de potasio 0,05M y pH 7.2 (BFP), [alpha] o [beta]-naftil acetato ([alpha] o [beta]NAF), o-dianisidina (OD), albumina serica bovina (ASB), homogeneizado de gorgojos (HM), dihidrocloruro de 3,3',5,5'-tetrametilbencidina (TMBZ), metanol (ME), acetato de sodio (ACE) 0,25M, pH 5, peroxido de hidrogeno 3% (PER), glutation reducido (GR), 1-cloro-2,4' dinitrobenzeno (
CDNB), yoduro de acetilcolina (ATCH), y 5,5'-ditiobis-2-acido nitrobenzoico (DTNB).
La actividad de glutation transferasa (GST, EC 2.5.1.18) fue determinada con 1-cloro-2-4dinitrobenceno (
CDNB) 1 mmol L -1 y GSH 1 mmol [[L.sup.-1]], siguiendo la formacion del complejo dinitrofenilglutation (e: 9,6 mmol [[L.sup.-1]] cm) a 340 nm en buffer fosfato de potasio 100 mmol [[L.sup.-1]] a pH 6,5.
Glutathione s-transferase activities were determined using the model substrates 1-chloro-2, 4-dinitrobenzene (
CDNB) and reduced GSH as substrates according to Habig et al (28) with slight modifications.
GST activity was measured using 1-chloro-2,4-dinitrobenzene (
CDNB) as a substrate by the method of Habig and Jacoby [26].
(f) [micro]mol of 1-chloro-2,4 dinitrobenzene (
CDNB)-glutathione(GSH) conjugated/min/mg protein.