Fig. 1. Acyl-cLIP measurement of palmitoylation of a SHH N-terminus peptide. (A) Schematic representation of the Acyl-cLIP assay for HHAT. (B) Polarisation of substrate and product SHH-FAM peptides at 1 μM and detergents at 2 mM or BSA at 0.15 mM; only BSA and detergents above their CMC increase polarised emission of the lipidated peptide. (C) DDM titration demonstrates specific Pal-SHH-FAM polarised emission above the CMC. (D) BSA titration demonstrates specific Pal-SHH-FAM polarised emission above Kd. (E) Real-time analysis of SHH-FAM palmitoylation showing HHAT-dependent increase in FA over time. Data represent mean ± SEM (assays performed in duplicate, n = 3).
