Our Zymogram Sample Buffer is supplied as ready to use solution and contains 62.5mM Tris, 4% SDS, 25% Glycerol and 0.01% Bromophenol Blue. It helps in solubilizing the proteins for loading onto the zymogram gels and provides bromophenol blue tracking dye to visualize the migration on zymogram gels.
Zymography is an electrophoretic technique, based on SDS-PAGE, that includes an enzyme substrate co-polymerized with polyacrylamide gel. Samples are prepared in Zymogram Sample Buffer without boiling to preserve the structure and activity of the enzyme. Following electrophoresis, the SDS is removed from the gel by washing in Zymogram Renature Buffer that contains a non-ionic detergent. The gels are then equilibrated in Zymogram Developing Buffer, which contains the divalent metal cation, required for enzymatic activity. The zymogram is subsequently stained (commonly with Amido Black or Coomassie Brilliant Blue), and areas of digestion appear as clear bands against a darkly stained background where the substrate has been degraded by the enzyme.
Features of Zymogram Sample Buffer:
• Ready to use Sample Buffer for protease analysis using zymogram gels
• Dry samples can be dissolved directly into the sample buffer