Open AccessIdentification of protease isozymes after analytical isoelectric focusing using fluorogenic substrates impregnated into cellulose membranes.
Author(s) -
Robert E. Smith
Publication year - 1984
Publication title -
journal of histochemistry and cytochemistry/the journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1177/32.12.6389692
Subject(s) - proteases , isoelectric focusing , membrane , protease , isozyme , chemistry , chromatography , cellulose , isoelectric point , substrate (aquarium) , biochemistry , enzyme , biology , ecology
A technique to characterize active isozymes of proteases is described. After isoelectric focusing in an ultrathin gel, a cellulose diacetate or hydrate membrane previously impregnated with media containing a fluorogenic substrate is applied to the surface of the gel. Low-molecular-weight peptide inhibitors can also be added to the membrane to enhance specificity. The technique shows excellent reproducibility, specificity, and sensitivity, and has promise for the study of isozymes of proteases in tissues and body fluids in normal and pathological states.