DOIAbstract
Methods to study protein S‐palmitoylation dynamics have previously relied on metabolic labeling with [14C]palmitate, which requires additional safety precautions and long exposures. Nonradioactive alkynyl palmitate analogs have been developed for in‐gel fluorescence detection and affinity purification. Cells metabolically labeled with the commercially available analog 17‐octadynoic acid are lysed and then combined with azide‐linked reporter tags for efficient conjugation by copper‐catalyzed click chemistry in phosphate buffer. This approach has been demonstrated to label hundreds of endogenous palmitoylated proteins and is compatible with traditional pulse‐chase methods. This protocol describes the reagents and procedures for labeling and d...
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