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Detection of Protease Activity by Fluorescent Peptide Zymography
JoVE Journal
Cancer Research
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JoVE Journal Cancer Research
Detection of Protease Activity by Fluorescent Peptide Zymography
DOI:

09:56 min

January 20, 2019

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Chapters

  • 00:04Title
  • 01:03Preparation of the Resolving Gel Layer
  • 01:52Preparation of the Azido-PEG3-maleimide Linker Molecule
  • 03:31Preparation of the Peptide Resolving Gel Layer
  • 05:16Preparation of the Stacking Gel
  • 06:05Electrophoresis of Biological Samples in Peptide Zymography Gels
  • 07:26Results: Detection of Protease Activity
  • 09:15Conclusion

Summary

Automatic Translation

Here, we present a detailed protocol for a modified zymographic technique in which fluorescent peptides are used as the degradable substrate in place of native proteins. Electrophoresis of biological samples in fluorescent peptide zymograms enables detection of a wider range of proteases than previous zymographic techniques.

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