ArticleSTAR protocols2025
Protocol for the visualization and identification of S-palmitoylated proteins in HCT-116 cells using metabolic labeling via click chemistry.
Article in STAR protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Reversible S-palmitoylation facilitates the regulation of a plethora of protein functions. Here, we present a protocol for enriching lipidated proteins with click-chemistry labeling and immunoprecipitation using biotin-azide-streptavidin interaction where protein activity assays are possible. We describe steps for performing the copper-based reaction for high-scale applications in milligram ranges, sample preparation, and the elution of palmitoylated-biotinylated proteins. Further, we detail procedures for the quantification of S-palmitoylation of isolated membrane proteins via reporter fluorophore on the western-blot level without lipid background. For complete details on the use and execution of this protocol, please refer to Merz et al.
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