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Methods Mol Biol. 2014;1120:33-41. doi: 10.1007/978-1-62703-791-4_3.

Metabolic labeling of Ras with tritiated palmitate to monitor palmitoylation and depalmitoylation.

Author information

1
Department of Medicine, NYU Langone School of Medicine, New York University, New York, NY, USA.

Abstract

Metabolic labeling with tritiated palmitate is a direct method for monitoring posttranslational modification of Ras proteins with this fatty acid. Advances in intensifying screens have allowed for the easy visualization of tritium without the need for extended exposure times. While more energetic radioisotopes are easier to visualize, the lack of commercial source and need for shielding make them more difficult to work with. Since radiolabeled palmitate is directly incorporated into Ras, its loss can be monitored by traditional pulse-chase experiments that cannot be accomplished with the method of acyl-exchange chemistry. As such, tritiated palmitate remains a readily accessible and direct method for monitoring the palmitoylation status of Ras proteins under a multitude of conditions.

PMID:
24470017
PMCID:
PMC4065800
DOI:
10.1007/978-1-62703-791-4_3
[Indexed for MEDLINE]
Free PMC Article

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