Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes. Anal Biochem 1984 Nov 15;143(1):30-4
Date
11/15/1984Pubmed ID
6442109DOI
10.1016/0003-2697(84)90553-0Scopus ID
2-s2.0-0021738226 (requires institutional sign-in at Scopus site) 511 CitationsAbstract
A simple inexpensive sensitive protease assay was developed using soluble fluorescein isothiocyanate (FITC)-labeled casein. Casein was reacted with FITC to form the fluorescein thiocarbamoyl derivative. This substrate is cleaved by trypsin, chymotrypsin, elastase, subtilisin, and thermolysin in a linear time-dependent manner. These enzymes can be measured in the nanogram and subnanogram range using this assay. The assay is reproducible, has a low blank, and uses casein, which resembles natural substrates of most proteases.
Author List
Twining SSAuthor
Sally S. Twining PhD Assistant Dean, Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCaseins
Chemical Phenomena
Chemistry
Fluorescein-5-isothiocyanate
Fluoresceins
Hydrogen-Ion Concentration
Hydrolysis
Macrophages
Peptide Hydrolases
Rabbits
Spectrometry, Fluorescence
Thiocyanates