Fluorescent assay of proteinases in cultured mammalian cells

Journal Article · Mon Jan 01 00:00:00 EST 1979 · J. Histochem. Cytochem.; (United States)

DOI:https://doi.org/10.1177/27.11.512330· OSTI ID:7099724

Dolbeare, F; Vanderlaan, M

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.

• Research Organization: Univ. of California, Livermore, CA (United States)
• OSTI ID: 7099724
• Journal Information: J. Histochem. Cytochem.; (United States), Vol. 27:11
• Country of Publication: United States
• Language: English

Related Subjects

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