D. Kohn, M. Weber, P. L. Carl
Sep 1, 1979
Citations
0
Influential Citations
3
Citations
Journal
Analytical biochemistry
Abstract
Abstract A peptidyl derivative of [ 3 H]aniline, Gly-Gly-Arg-[ 3 H]anilide, can be used as a substrate in a convenient and sensitive assay procedure for trypsin, urokinase, and plasminogen activator from transformed cells. The extent of hydrolysis can be determined simply by selective extraction of the product [ 3 H]aniline into an organic phase containing a scintillant. (The uncleaved peptide is not appreciably soluble in this phase and is not counted.) The reaction is of comparable sensitivity to fluorimetric assays, but has the advantage that no cleanup of the biological sample is required, since it is far less subject to interference from fluorescence quenching. Other peptidyl anilides should be useful for assaying proteolytic enzymes with widely varying specificities.