Assays

Phosphoproteomics: starting from the protein extracts from the global proteomics experiments, proteases inhibitors (Sigma, P8340) and phosphatases inhibitors (final concentration in Na3VO4 = 1 mM) were added to all samples. Protein concentration was determined using RC-DC assay (BioRad, Hercules, CA, USA) according to the manufacturer’s instructions. 250 µg of proteins for each sample were reduced and alkylated prior to an in-house optimized single-pot, solid-phase-enhanced sample preparation ...