Nitro blue tetrazolium

Often these methods are not sensitive enough for quantitation of different CK isoenzymes. Konttinen and Somer (45) used electrophoresis on agarose to obtain sharper bands and increased sensitivity. Staining with nitro-blue tetrazolium was found to... 

NAP-I, NAP-2 Neutrophilactivating peptides -1 and -2 NBT Nitro-blue tetrazolium NCI Non-collagen 1 N-CAM Neural cell adhesion molecule... 

Buffer and chemicals pH 7.8 phosphate buffer (16.29g Na2HP04-2H20, 1.17g NaH2P04H20, 1000 ml distilled water), Ethylene Diamine Tetra Acetic Acid (EDTA), Methionine (Met), Polyvinylpyrrolidone (PVP), Triton X-100, Phenylmethylsulphonylfluoride (PMSF), Riboflavin, Nitro Blue Tetrazolium (NBT). 

The on-bead assay was conducted according to Scheme 3.19, which shows the chain of events, which leads to a colorimetric response, when an oligosaccharide binds effectively to the B. purpurea lectin. The lectin was covalently linked to biotin, a small molecule with an extremely high affinity for streptavidin. The bead-lectin-biotin conjugates were then exposed to streptavidin, linked to the enzyme alkaline phosphatase. Alkaline phosphatase hydrolyses phosphate esters [e.g., 5-bromo-4-chloro-3-indolyl phosphate (BCIP), 110]. When the 5-bromo-4-chloro-3-hydroxyindole (111) is released, in the presence of nitro blue tetrazolium (NBT), it forms a dark purple, insoluble dye, thus staining beads where there was a favorable binding interaction. 

Borregaard, N., Cross, A. R., Herlin, T., Jones, O. T. G., Segal, A. W., Valerius, N. H. (1983). A variant form of X-linked chronic granulomatous disease with normal nitro-blue tetrazolium slide test and cytochrome b. Eur. J. Clin. Invest. 13,243-7. 

Bromochloroindolyl Phosphate-Nitro blue Tetrazolium (BCIP/NBT) EDTA Purple Black/NA... 

List of Abbreviations BCIP, 5-bromo-4-chloro-3-indolyl phosphate, toluidine salt CCD, charged-coupled device CNS, central nervous system cDNA, copy or complimentary RNA cRNA, copy or complimentary RNA DNase, deoxyribonuclease FITC, fluorescein isothiocyanate mRNA, messenger RNA NBT, nitro blue tetrazolium choride PCR, polymerase chain reaction RNase, ribonclease... 

Alkaline phosphatase acts on many substrates as well, each precipitating as a different color. For example, a combination of 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium (NBT) results in a permanent blue precipitate at the site of alkaline phosphatase localization. There are other compounds that can also be tried, such as Fast Red TR/Naphthol AS-MX (Sigma, St. Louis, MO), which precipitates as a red color. 

Prepare substtate solution 10.0 mL buffer 2, 27.7 pL Nitro-blue tetrazolium (NBT), 22.5 pL BCIP, 6.0 drops Levamisole (1 mM). 

The evidence that O is a product of the burst has already been summarized. Another direct consequence of the formation of O is the reduction of nitroblue tetrazolium to form an insoluble deposit of blue formazan. The reduction of nitro blue tetrazolium had been observed to occur in PMNs in patients with bacterial infections. In intact cells this appears to be a consequence of the formation of O it fails to occur in PMNs from patients with chronic granulomatous disease which cannot undergo the respiratory burst it is diminished by anaerobiosis and it is inhibitable by superoxide dismutase... 

It is convenient to have a method for the detection and rough estimation of urease activity in gels. A simple staining procedure using cresol red has been described (27), but a more elaborate procedure (28) employing p-nitro blue tetrazolium results in sharper definition. 

Similarly, nitro blue tetrazolium is reduced by 02 (k = 3 x 104 dm3 mol 1 s 1), and the mono-reduced species subsequently disproportionates yielding the two-electron-reduced monoformazan which absorbs in the visible (e(530) = 2.34 x 103 dm3 mol 1 cm"1 Bielski et al. 1980 for some of the problems that one may encounter using this assay see Cabelli 1997]. 

Bielski BFIJ, Shiue GG, Bajuk S (1980) Reduction of nitro blue tetrazolium by C02 and O2 radicals. J Phys Chem 84 830-833... 

The SOD activity measurements using the xanthine-xanthine oxidase nitro blue tetrazolium (NBT) method [45] indicated that a concentration of 0.75 xM of [Mn(OBz)(H-3,5-/-Pr2pz)(L 1)] 1 and 0.8 iM of the related [Mn(OBz)(Ll)] 2 complex exert the same SOD activity as one unit of native SOD. The high SOD activity of the tris(pyrazolyl)borate complexes was attributed to their structural similarities to the MnSOD active site [45],... 

Additional substrates include naphthol AS-BI phosphate, naphthol AS-TR phosphate and 5-bromo-4-chloro-3-indoxyl phosphate (BCIP). Other possible chromogens include Fast Red LB, Fast Garnet GBC, Nitro Blue Tetrazolium (NBT) and lodonitrotetrazolium Violet (INT). 

Prepare the color development solution by adding the BCIP [25 mg of 5-bromo-4-chloro-3-indolyl phosphate, p-toluidine salt (Sigma Chemical Co.) in 2.5 mL of N,N -dimethylformamide] and NBT (50 mg of nitro blue tetrazolium in 5 mL of 50% N, N -dimethylformamide) solutions to 100 mL of 1.0 M Tris-HCl, pH 9.0. 

The polyphenol extract from the barley bran (BPE) induced the differentiation of human promyelocytic leukemia HL-60 cells, based on the expression of the differentiation markers such as nitro blue tetrazolium (NBT) reducing activity and a-naphthyl butyrate esterase activity. Prodelphini-din B-3, Tl, T2, and T3 induced 26-40% NBT-positive cells and 22-32% a-naphthyl butyrate esterase-positive cells. Proanthocyanidins might potentiate the retinoic acid (all-frarcs-retinoic acid)-induced granulocytic differentiation and sodium butyrate-induced monocytic differentiation in the HL-60 cells [101]. 

The assistance of several postdoctoral and doctoral fellows must be acknowledged in particular, Dr. Patrick McCarthy for his major contribution to the chapter on techniques for measuring lipid peroxidation Dr. Parves Haris for his assistance and expertise in the preparation of the section on Fourier Transform Infra Red Spectroscopy, Mr. George Paganga for the section on the preparation of low density lipoproteins and Dr. Paul Eggleton for the preparation of the section on the nitro blue tetrazolium assay. 

Stain for cytotoxicity based on change to formazan on reduction. See nitro blue tetrazolium, which has similar chemistry and higher use. 

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