DNSA (1-Dimethylamino-naphthelene-5-sulfonamide) forms with carbonic anhydrase (carbonate hydrolase, EC 4.2.1.1) a highly fluorescent complex [1]. We used the formation of these complexes for the detection of the isoenzyme bands of carbonic anhydrase in isoelectric focusing and electrophoresis in acrylamide gels. DNSA is added to the solution of the isoenzymes before separation. Using quartz tubes - the maximum of the excitation spectrum is 280 nm [1] - for supporting the gels the fluorescent enzyme bands can be observed during development in UV light and without putting the gels out of the quartz tubes. This technique allows the detection of the carbonic anhydrase isoenzymes with greater precision and simplicity than with any staining method.
Holke
M
M
Zentralinstitut für Biochemie und Biophysik der Freien Universität Berlin, 1 33, Berlin, Germany
Siegmund
P
P
Detektion von carboanhydrase isoenzymen in acrylamidgelen mit einem fluoreszierenden sulfonamid.