Membrane targeting of palmitoylated Wnt and Hedgehog revealed by chemical probes

► We describe a method for metabolic labeling of palmitoylated Wnt and Hh. ► We use w-alkynyl palmitic acid and Click chemistry. ► The labeling does not interfere with Wnt or Hh signaling. ► Palmitoylated Wnt and Hh proteins are found in cellular membrane fractions. ► The method is useful for validating inhibitors of Wnt and Hh acyltransferases.