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In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening
JoVE Journal
Bioengenharia
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JoVE Journal Bioengenharia
In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening

In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening

DOI:
10.3791/64392-v

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08:16 min

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October 06, 2022

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1Department of Molecular, Cell and Cancer Biology,UMass Chan Medical School

Capítulos

  • 00:04Introduction
  • 01:15Small‐Scale Recombinant Caspases Extraction
  • 02:34Small‐Scale His‐Tagged Caspases Purification
  • 04:09In Vitro Cleavage Assay with Substrates Generated with RRL
  • 05:29Results: In Vitro Cleavage Assays to Express and Purify Recombinant Drosophila Caspases
  • 07:33Conclusion

Summary

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Tadução automática

Here we present a protocol to express and purify recombinant Drosophila caspases Dronc and Drice, and their use in in vitro cleavage assays.

Tags

In Vitro Cleavage AssayCaspasesDrosophilaDroncRecombinant ProteinsSubstrate ScreeningCell LysisNi-NTA AgarosePurificationEnzymatic Activity

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