Blood was mixed with EDTA (1:1 ratio) prior to its incubation with different charged NPs for 15 mins at 37[degrees]C in the presence of nitro blue tetrazolium
Superoxide radical ([O.sub.2.sup..-]) 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., 2015; Marioni et al., 2016).
Abbreviations CMCS: Cultured mycelium Cordyceps sinensis C[Cl.sub.4]: Carbon tetrachloride ALT: Alanine aminotransferase AST: Aspartate aminotransferase Alb: Albumin T.Bil: Total bilirubin Hyp: Hydroxyproline H&E: Hematoxylin and eosin [alpha]-SMA: Alpha smooth muscle actin HPLC: High performance liquid chromatography CCK8: Cell Counting Kit-8 MTT: Thiazolyl Blue Tetrazolium
Blue HSCs: Hepatic stellate cells DMEM: Dulbecco's modified Eagle's medium FBS: Fetal bovine serum DMSO: Dimethyl sulfoxide BSA: Bovine plasma albumin PBS: Phosphate-buffer saline DAB: 3,3'-Diamino Benzedrine DAPI: 4',6-Diamidino-2-phenylindole TGF-[beta]: Transforming growth factor-beta BW: Body weight.
It was assayed by the reduction of nitro blue tetrazolium
Under alkaline conditions, fructosamine reverts to the eneaminol form, which reduces nitro blue tetrazolium
to a formazan that can be quantified by measuring the absorbance (17).
Blots were stained for alkaline phosphatase using 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium
(NBT) (Jilani et al., 1996; Lamelli, 1970; Towbin et al., 1979).
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 6.000)a em triplicata.
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.sub.2] in 100 mM Tris-HCl, pH 9.5.