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Cathepsin B2 measurement by sensitive fluorometric ammonia analysis

✍ Scribed by S. Taylor; V. Ninjoor; D.M. Dowd; A.L. Tappel

Book ID: 102984163
Publisher: Elsevier Science
Year: 1974
Tongue: English
Weight: 579 KB
Volume: 60
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(74)90140-7

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✦ Synopsis

A procedure has been developed for the measurement of cathepsin B2 activity that is baaed upon highly sensitive fluorometric ammonia analysis. The fluorochrome results from the reaction of phthaldehyde and mercaptoethanol with ammonium salts at pH 7.4. The method is sensitive in the range of 5400 nmoles ammonium salt/ml. The assay is highly specific for ammonia since amino acids, peptides, amines, and amides do not interfere. Likewise, other components of the cathepsin B2 reaction system do not interfere. The most distinct advantage of the method is that the awkward diffusion step of most routine ammonia analyses can be eliminated.

Methods

Enzymes. The source of cathepsin B2 activity was a rat liver homogenate. The homogenate was prepared in a Waring Blendor with 10 vol 153

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