Zymography, the detection, identification, and even quantification of enzyme activity fractionated by gel electrophoresis, has received increasing attention in the last years, as revealed by the number of articles published. A number of enzymes are routinely detected by zymography, especially with clinical interest. This introductory chapter reviews the major principles behind zymography. New advances of this method are basically focused towards two-dimensional zymography and transfer zymography as will be explained in the rest of the chapters. Some general considerations when performing the experiments are outlined as well as the major troubleshooting and safety issues necessary for correct development of the electrophoresis.
Zymography, the detection of enzymatic activity on gel electrophoresis, has been a technique described in the literature for at least in the past 50 years. Although a diverse amount of enzymes, especially proteases, have been detected, advances and improvements have been slower in comparison with other molecular biology, biotechnology and chromatography techniques. Most of the reviews and patents published focus on the technique as an element for enzymatic testing, but detailed analytical studies are scarce. Patents referring to zymography per se are few and the technique itself is hardly an important issue in titles or keywords in many scientific publications. This review covers a small condensation of the works published so far dealing with the identification of proteolytic enzymes in electrophoretic gel supports and its variations like 2-D zymography, real-time zymography, and in-situ zymography. Moreover, a scope will be given to visualize the new tendencies of this method, regarding substrates used and activity visualization. What to expect from zymography in the near future is also approached.
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