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Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins
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Biochemistry
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JoVE Journal Biochemistry
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:
10.3791/61018-v

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08:28 min

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March 29, 2020

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,
1Department of Pharmacology,UC Davis School of Medicine

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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Automatic Translation

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.

Tags

PalmitoylationBrain Membrane ProteinsAcyl-PEGyl ExchangeGel Shift AssayQuantitative DeterminationBiochemical MethodAffinity PurificationGel MobilityHomogenizationCentrifugationABCA AssayProtein QuantitationTCEPNEMChloroform-methanol Precipitation

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