Medical College of Wisconsin
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Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes. Anal Biochem 1984 Nov 15;143(1):30-4

Date

11/15/1984

Pubmed ID

6442109

DOI

10.1016/0003-2697(84)90553-0

Scopus ID

2-s2.0-0021738226 (requires institutional sign-in at Scopus site)   533 Citations

Abstract

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 SS

Author

Sally S. Twining PhD Assistant Dean, Professor in the Biochemistry department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Caseins
Chemical Phenomena
Chemistry
Fluorescein-5-isothiocyanate
Fluoresceins
Hydrogen-Ion Concentration
Hydrolysis
Macrophages
Peptide Hydrolases
Rabbits
Spectrometry, Fluorescence
Thiocyanates