Zymography is an electrophoretic technique that includes a substrate copolymerised with the polyacrylamide gel for the detection of enzymes and their activity. Samples are prepared without denaturing the active enzymes present in the samples. Following electrophoresis, the gel is placed in an enzyme activation buffer which allows the enzymes present in the sample to become active and digest the substrates copolymerised in the gel. The zymogram is subsequently stained and the areas of enzyme activity and digestion become visible.
Gelatin is the most commonly used substrate, and is useful for demonstrating the activity of gelatin-degrading proteases, but zymography has been applied to a variety of enzymes, including xylanases, proteases, lipases, etc. Suitable for drug discovery and screening of low abundant enzymes and their isomers in complex mixtures.
Applications -This lab activity teaches the following:
- A modified protein electrophoresis technique.
- Use of electrophoresis to study enzyme activity.
- Introduction to zymograms.
Please refer to the Product Protocol for additional equipment needed.Click here to download Product Protocol.