was detected by their oxidative action that causes the nitro blue tetrazolium
(100 [micro]l of NBT 1 mg/ml, Sigma-Aldrich) reduction to nitroblue diformazan, by following the methodology previously described by us (Peralta et al.
Blots were stained for alkaline phosphatase using 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium
(NBT) (Jilani et al.
After 48 hours of incubation at 37[degrees]C, 50 [micro]l thiazolyl blue tetrazolium
bromide (MTT) (5 mg/ml in PBS) reagent was added to each well.
Chemicals used in this study were 2, 2-diphenyl-1-picrylhydrazyl (DPPH), potassium ferricyanide, sodium nitrite, trichloroacetic acid, Folin-Ciocalteu reagent, sodium nitroprusside, sulfanilamide, N-1-naphthylethylenediamine dihydrochloride, butylated hydroxy anisole (BHA), ascorbic acid, [alpha]-tocopheryl acetate, ethylenediamine tetra-acetic acid, phosphoric acid, naphthylethylenediamine dihydrochloride, nitro blue tetrazolium
, NADH, phenazine methosulfate, ferrous ammonium sulfate, DMSO and hydrogen peroxide.
Blood samples were obtained from the caudal vein of six fish from each tank (18 fish per treatment) using heparinized syringes after anesthetizing the fish with tricaine methanesulfonate (MS-222) at a concentration of 100 mg/L for hematocrit, hemoglobin, nitro blue tetrazolium
(NBT) activity, and red blood and white blood cell counts.
Previamente aos ensaios com os virus, foram realizados testes de citotoxicidade (teste de MTT, Thiazolyl Blue Tetrazolium
Bromide), avaliando-se a viabilidade de uma quantidade constante de celulas MDBK (1,6x105 por cavidade da placa de 96 pocos) frente a diferentes concentracoes de PEG (PM 6.
Thiazol blue tetrazolium
was added (10 [micro]L) and the plate was returned to thermostat for 5 h.
Color development was presented in 20 mL of a mixture consisting of 7 mg nitro blue tetrazolium
, 5 mg 5-bromo-4-chloro-3-indolyl-phosphate, 100 mM NaCl, and 5 mM Mg[Cl.
Superoxide dismutase activity was determined by measuring its ability to inhibit the photochemical reduction of nitro blue tetrazolium
chloride (NBT, 2,2'-di-p-nitrophenyl-5,5'-diphenyl-[3,3-dimethoxy-4,4'-diphenylene] ditetrazolium chloride) according to the method of Giannopolitis and Ries (1977), with slight modifications.
Probes were visualized by bright field microscopy following formation of an oligoDIG-antiDIG-alkaline phosphatase conjugate treated with 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium
The filters were then washed and stained with streptavidin-linked alkaline phosphatase and nitro blue tetrazolium
salt, using Blugene (Bethesda Research Laboratories) reagents and protocols.
Thiazolyl blue tetrazolium
bromide (MTT), galantamine hydrobromide and AChE from Electrophorus electricus (electric eel), 500U/mg, were purchased from Sigma (St.