Casein zymography is a technique used to detect casein-degrading enzymes in biological samples.
Procedure
- Preparation: A polyacrylamide gel is co-polymerized with casein as the substrate.
- Electrophoresis: Samples containing proteases (e.g., conditioned media or cell lysates) are separated under non-reducing, denaturing conditions.
- Renaturation & Incubation: After electrophoresis, SDS is removed, and the gel is incubated in a buffer that promotes enzyme renaturation and activity.
- Staining: The gel is stained with Coomassie Blue, and clear bands appear where casein has been degraded, indicating caseinolytic activity.
