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Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins

Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins

DOI:

08:28 min

March 29, 2020

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Chapters

  • 00:04Introduction
  • 00:40Preparation of Mouse Brain Membranes
  • 02:29Acyl-PEGyl Exchange Gel-Shift (APEGS) Assay
  • 06:54Western Blot Analysis of PEGylated Proteins
  • 07:11Results: Palmitoylation of SynDIG Proteins
  • 07:41Conclusion

Summary

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Palmitoylation entails the incorporation of a 16-carbon palmitate moiety to cysteine residues of target proteins in a reversible manner. Here, we describe a biochemical approach, the acyl-PEGyl exchange gel shift (APEGS) assay, to investigate the palmitoylation state of any protein of interest in mouse brain lysates.

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