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Title: Fluorescent assay of proteinases in cultured mammalian cells

Abstract

We have demonstrated proteinase activity in unfixed cells grown on tissue culture plates with a technique using 5-nitrosalicylaldehyde and peptide derivatives of 4-methoxy-2-naphthylamine. The 4-methoxy-2-naphthylamine liberated by proteinase activity reacts with 5-nitrosalicylaldehyde to form a fluorescent product. The substrates CBZ-alanyl-arginyl-arginyl-4-methoxy-2-naphthylamine and lysyl-alanyl-4-methoxy-2-naphthylamine, were used for the direct visual detection of two arylamidase activities in BALB/c 3T3 and C3H 10T1/2 cells. With low magnification these enzyme activities can be detected in single clones; with higher magnification the fluorescent product can be seen within the cytoplasm of single cells.

Authors:
;
Publication Date:
Research Org.:
Univ. of California, Livermore
OSTI Identifier:
7099724
Resource Type:
Journal Article
Journal Name:
J. Histochem. Cytochem.; (United States)
Additional Journal Information:
Journal Volume: 27:11
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ENZYME ACTIVITY; BIOASSAY; ENZYMES; BIOCHEMICAL REACTION KINETICS; CELL CULTURES; CULTURE MEDIA; FLUORESCENCE; METABOLISM; MICE; ANIMALS; KINETICS; LUMINESCENCE; MAMMALS; REACTION KINETICS; RODENTS; VERTEBRATES; 550200* - Biochemistry; 550300 - Cytology

Citation Formats

Dolbeare, F, and Vanderlaan, M. Fluorescent assay of proteinases in cultured mammalian cells. United States: N. p., 1979. Web. doi:10.1177/27.11.512330.
Dolbeare, F, & Vanderlaan, M. Fluorescent assay of proteinases in cultured mammalian cells. United States. https://doi.org/10.1177/27.11.512330
Dolbeare, F, and Vanderlaan, M. Mon . "Fluorescent assay of proteinases in cultured mammalian cells". United States. https://doi.org/10.1177/27.11.512330.
@article{osti_7099724,
title = {Fluorescent assay of proteinases in cultured mammalian cells},
author = {Dolbeare, F and Vanderlaan, M},
abstractNote = {We have demonstrated proteinase activity in unfixed cells grown on tissue culture plates with a technique using 5-nitrosalicylaldehyde and peptide derivatives of 4-methoxy-2-naphthylamine. The 4-methoxy-2-naphthylamine liberated by proteinase activity reacts with 5-nitrosalicylaldehyde to form a fluorescent product. The substrates CBZ-alanyl-arginyl-arginyl-4-methoxy-2-naphthylamine and lysyl-alanyl-4-methoxy-2-naphthylamine, were used for the direct visual detection of two arylamidase activities in BALB/c 3T3 and C3H 10T1/2 cells. With low magnification these enzyme activities can be detected in single clones; with higher magnification the fluorescent product can be seen within the cytoplasm of single cells.},
doi = {10.1177/27.11.512330},
url = {https://www.osti.gov/biblio/7099724}, journal = {J. Histochem. Cytochem.; (United States)},
number = ,
volume = 27:11,
place = {United States},
year = {1979},
month = {1}
}