Preparation of Nessler's Reagent

Vanselow, A. P.

doi:10.1021/ac50149a007

Cited by 101 publications

(31 citation statements)

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“…The tissue was extracted three times with about 100 volumes of chloroform: methanol as was done for the preparation of mixed brain phospholipids. An aliquot containing [12][13][14][15][16][17][18][19][20] 1jg P was applied to silica gel plates (Camag DO, 400 gA, 20 X 20 cm) and chromatographed in two dimensions. The solvents used were chloroform: methanol: acetic acid: water (25: 15: 4: 2) (14) in the first dimension, and chloroform: acetone: methanol: acetic acid: water (5: 2: 1: 1: 0.5) (10) in the second.…”

Section: Methodsmentioning

confidence: 99%

The phospholipid requirement of tissue factor in blood coagulation

Nemerson¹

1968

J. Clin. Invest.

142

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Section: Methodsmentioning

confidence: 99%

The phospholipid requirement of tissue factor in blood coagulation

Nemerson¹

1968

J. Clin. Invest.

142

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“…The following compounds were tested for substrate speci- The release of NH3 from acetamide, propionamide and IIbutyramide was determined at 436 nm using Nessler's reagent [13]. The activity for other compounds was measured by following the increase in the products by HPLC using the following solvent systems; ( 3 ) for aniline, p-toluidine, maminophenol, p-aminophenol, rn-phenylenediamine, p-phenylenediamine, 4-hydroxy-o-toluidine, and N-acetyl-L-tyrosine, 73 mM KH2P04 (pH 5.…”

Section: Substrate Specificitymentioning

confidence: 99%

Purification and characterization of aryl acylamidase from Nocardia globerula

Yoshioka

Nagasawa

Yamada

1991

European Journal of Biochemistry

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Aryl acylamidase was purified from an extract of N-acetyl-o-toluidine-induced cells of Nocardiu globerulu I F 0 13510 in ten steps. The purified enzyme appeared to be homogeneous from analysis by polyacrylamide gel electrophoresis. The enzyme has a molecular mass of approximately 126 kDa and consists of two subunits which are identical in molecular mass. The purified enzyme catalyzed the hydrolysis of N-acetyl-o-toluidine to o-toluidine and acetic acid at a rate of 47.7 pmol . min-' . mg-' at 35°C. It also catalyzed the hydrolysis of various anilide derivatives and esters, as well as the transfer of an acetyl group to aniline as an acetyl acceptor. The purified enzyme was sensitive to thiol reagents such as HgClz and p-chloromercuribenzoate. The amino-terminal sequence (28 amino acid residues) of the enzyme was determined. Based on the substrate specificity of this enzyme, the pathway intermediates involved in the conversion of n-acetyl-o-toluidine to 4-hydroxy-N-acetyl-o-toluidine are discussed Microbial hydroxylation of aromatic compounds catalyzes position-specific and stereospecific reactions efficiently. This reaction has been utilized in efforts to produce useful compounds in the field of medicine and for chemical industries 6wc0cH3 -t 1/2 0, -no &c0cH3 ( 1 1 Recently, we surveyed the hydroxylation of N-acetyl-otoluidine to 4'-hydroxy-N-acetyl-o-toluidine by various bacteria and actinomycetes. Nocardia globerula I F 0 13510 was selected as the best strain to accumulate 4-hydroxy-N-acetylo-toluidine [lo]. As soon as N-acetyl-o-toluidine was added to culture broth of N. globerulu, o-toluidine was formed. Subsequently, in accordance with the decrease in o-toluidine, an accumulation of 4'-hydroxy-N-acetyl-o-toluidine in the culture broth was observed. Therefore, we presumed that the first step may be the hydrolysis of N-acetyl-o-toluidine to o-toluidine by aryl acylamidase, being associated with the conversion of N-acetyl-o-toluidine into 4-hydroxy-N-acetylo-toluidine.In the present study, in order to elucidate the accumulation mechanism of 4-hydroxy-N-acetyl-o-toluidine, we attemptedCorrespondence to

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“…The specific activity curve of urinary urea was determined on urea isolated by xanthydrol precipitation (6). Urea excretion was determined by the urease method followed by Nesslerization (7). The "urea pool" was measured on two occasions (Studies 1 and 4) by body water determina-The problem of understanding the model and its limitations is that of relating the mathematics to biological meaning.…”

Section: Methodsmentioning

confidence: 99%