NITRO BLUE TETRAZOLIUM NBT

Merck Index Number Not listed Chemical/Dye Class Tetrazolium salt Moleeular Formula C40H30CI2N10O6 Molecular Weight 817.64 

CA Index Name 2H-Tetrazolium, 2,2 -(3,3 -dimethoxy [l,l -biphenyl]-4,4 -diyl)bis[3-(4-nitrophenyl)-5-phe-nyl]-, chloride (1 2) 

Other Names 2//-Tetrazolium, 2,2 -(3,3 -dimethoxy [l,l -biphenyl]-4,4 -diyl)bis[3-(4-nitrophenyl)-5-phe-nyl]-, dichloride 27/-Tetrazolium, 3,3 -(3,3 -dimethoxy-4,4 -biphenylylene)bis[2-(p-nitrophenyl)-5-phenyl]-, dichloride Tetrazolium, 3,3 -(3,3 -dimethoxy-4,4 -biphe-nylylene)bis[2-(p-nitrophenyl)-5-phenyl-2H-chloride] 2, 2 -Bis(p-nitrophenyl)-5,5 -diphenyl-3,3 -(3,3 -dimethoxy-4,4 -biphenylylene)ditetrazolium chloride 2,2 -Bis(p-ni-trophenyl)-5,5 -diphenyl-3,3 -(3,3 -dimethoxy-4,4 -di-phenylene)ditetrazolium chloride 2,2 -Di-p-nitrophenyl-5,5 -diphenyl-3,3 -bis(3,3 -dimethoxy-4,4 -biphenylene) ditetrazolium chloride 2,2 -Dinitrophenyl-5,5 -diphenyl-3,3 -dimethoxy-4,4 -diphenylene)ditetrazolium chloride 3,3 -(3,3 -Dimethoxy-4,4 -biphenylylene)bis[2-(p-nitro-phenyl)-5-phenyl-2H-tetrazolium chloride] 3,3 -(3,3 -Dimethoxy-4,4 -diphenylene)bis[2-(p-nitrophenyl)-5-phenyltetrazolium chloride] NBT NBT (dye) NSC 27622 NTB Nitro BT Nitro Blue Tetrazolium Nitro Blue Tetrazolium chloride Nitro Tetrazolium BT Nitro-tetrazolium Chloride Blue Nitrotetrazolium blue Tetrazolium Nitro BT Tetrazolium nitro blue /)-NBT / -Nitro blue tetrazolium /)-Nitro blue tetrazolium chloride p-Nitrotetrazolium blue 

Physical Form Yellow powder Solubility Soluble in water, ethanol, methanol, dimethyl sulfoxide 

Melting Point 189 °C Absorption ( ax) 256 nm Synthesis Synthetic methods  

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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. 

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 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). 

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]. 

Nitro blue tetrazolium (NBT Fisher Scientific, Fair Lawn, New Jersey). 

Hearts are weighed, sectioned transversely (5 or 6 slices), and stained for 20 min in 0.05% nitro blue tetrazolium (NBT Fisher Scientific, Fair Lawn, New Jersey) at 60°C (18, 19, 25). 

Reports dealing with mechanistic studies of NOS indicate that the enzyme can transfer electrons from NADPH to molecular oxygen via FAD or FMN to form superoxide anion and H2O2 in the presence of calcium or calmodulin (Mayer et al., 1990 Pou etal., 1992). One of these studies also demonstrated the concentration-dependent diminution of superoxide anion generation in the presence of N -nitro-L-arginine methyl ester (l-NAME). NOS has also been shown to donate electrons to acceptors, including nitro-blue tetrazolium (NBT) (Dawson et al., 1991) and cytochrome c (Klatt et al., 1992). These studies have used L-citrulhne rather than NO determination to detect NOS activity. 

BRL DNA Detection kit (no. 530-8239SA). This kit contains 120 jL BRL streptavidm, 60 pL biotinylated calf intestinal alkaline phosphatase, 660 pL nitro-blue tetrazolium (NBT), 500 pL 5-bromo-4-chloro-3-indolyl phosphate (BCIP), 25 pg/pL biotinylated DNA. 

Probes prepared with either digoxigenin- or biotin-modified nucleotides can be hybridized to Southern blots to detect target nucleic acid sequences. These methods offer an attractive alternative to radioactively tagged probes in terms of safety, cost, and efficiency. Most previous nonradioactive strategies utilized the detection of the modified base by the use of a coupled antibody- or avidin-alkaline phosphatase. The blot with the bound alkaline phosphatase was then treated with a compound, such as Nitro Blue Tetrazolium (NBT), that was converted to an insoluble, colored compound at the site of hybridization, thus facilitating visualization of the hybridized probe. 

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