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Detection of Protein S-Acylation using Acyl-Resin Assisted Capture
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Biochemistry
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JoVE Journal Biochemistry
Detection of Protein S-Acylation using Acyl-Resin Assisted Capture

Detection of Protein S-Acylation using Acyl-Resin Assisted Capture

DOI:
10.3791/61016-v

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08:31 min

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April 10, 2020

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, , ,
1Department of Biochemistry and Molecular Biology,McGovern Medical School at UT Health, 2MD Anderson UT Health Graduate School

Chapters

  • 00:04Introduction
  • 01:25Cell Lysate Preparation
  • 02:26Acyl-Resin Assisted Capture (Acyl-RAC): Free Thiol Group Blocking
  • 03:54Acyl-RAC: Hydroxylamine (HAM) Cleavage and S-Acylated Protein Capture
  • 05:52S-Acylated Protein Elution and Detection
  • 06:52Results: Representative S-Acylated Protein Detection
  • 07:43Conclusion

Summary

Automatic Translation

Automatic Translation

Acyl-RAC (Acyl-Resin Assisted Capture) is a highly sensitive, reliable and easy to perform method to detect reversible lipid modification of cysteine residues (S-acylation) in a variety of biological samples.

Tags

Keywords: Acyl-resin Assisted CaptureAcyl-RACProtein S-acylationThioester Bond CleavageChloroform-methanol PrecipitationProtein PelletCell LysisProtein Concentration EstimationBradford AssayBCA Assay

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