(a) Succinate dehydrogenase (SDH) activity in liver protein extracts of ApoE KO mice was estimated with colorimetric method containing nitroblue tetrazolium
2 mL of clotted blood and 1 mL of fresh EDTA anticoagulated blood were collected for serum alpha-1 AT assay and nitroblue tetrazolium
Laboratory evaluation for neutrophil's oxidative burst was performed and diagnosis of chronic granulomatous disease (CGD) was confirmed by both negative Nitroblue tetrazolium
test (NBT) and lack of dihydrorhodamine (DHR) fluorescence following neutrophil stimulation against phorbol myristate acetate (PMA).
Chemical reagents including a JNK inhibitor (JNKI), brefeldin A (BFA), SP600125 (SP), BCIP, MTT, and nitroblue tetrazolium
(NBT) were obtained from Sigma Chemical (St.
Fructosamine test was conducted using nitroblue tetrazolium
Immunologic studies including serum IgG IgA and IgM and phagocytic function tested with nitroblue tetrazolium
were within normal limits.
2,6-Dichloro-indophenol (DFI), Folin-Ciocalteu Reagent, Formic acid, Acetonitrile, Anthrone, Cyanidin 3-rhamnoside, Quercetin 3-rhamnoside, Pelargonidin 3-rhamnoside, 2,2-Azinobis-3-ethylbenzthiazoline-6-sulphonic acid radical cation ([ABTS.sup.*+]), 6-Hydroxy-2,5,7,8tetramethylchroman-2-carboxylic acid (Trolox), Ethylenediaminetetracetic acid (EDTA), Nitroblue tetrazolium
chloride (NBT) and Hydrogen peroxide ([H.sub.2][O.sub.2]) were purchased from Sigma Chemical Co.
Activity was determined by measuring its ability to inhibit the photochemical reduction of nitroblue tetrazolium
Para evaluar la respuesta inmune se utilizo el metodo de reduccion de Nitroblue Tetrazolium
(NBT) por ensayo espectrofotometrico descrito en Mohanty & Sahoo (41).
Pick, "Microassays for superoxide and hydrogen peroxide production and nitroblue tetrazolium
reduction using an enzyme immunoassay microplate reader," Methods in Enzymology, vol.
Abbreviations BSA: Bovine serum albumin PAGE: Polyacrylamide gel electrophoresis NBT: Nitroblue tetrazolium
PMS: Phenazine methosulphate PPP: Pentose phosphate pathway [NADP.sup.+]: [beta]-Nicotinamide adenine dinucleotide phosphate NADPH: Reduced [beta]-Nicotinamide adenine dinucleotide phosphate G6PD: Glucose-6-phosphate dehydrogenase.
Finally, the sections were kept in the dark at room temperature in the colour detection substrate solution nitroblue tetrazolium
and 5-bromo-4-chloro-3'-Indolylphosphate (BCIP/NBT) as recommended by manufacturer' (Roche) in NTM until appearance of the colour.