Target protein identification of SB2301 by proteomics study

Target protein identification study of SB2301, a small molecule that reduces cellular lipid droplets, was performed using the thermal stability shift-based fluorescence difference in two-dimensional gel electrophoresis (TS-FITGE) The protein spots from the silver-stained gel were excised, destained, and digested with trypsin. The mixture was evaporated in SpeedVac and then dissolved in 10% acetonitrile with 0.1% formic acid. The resulting peptides were desalted in a trap column (180 μm × 20 mm, Symmetry C18) and separated on a C18 reversed-phase analytical column (75 μm × 200 mm, 1.7 μm, BEH130 C18) (Waters) with an electrospray ionization Pico Tip (±10 μm i.d.) [New objective]. The data were converted to .pkl files by Protein Lynx Global Server and searched by MASCOT engine with the SwissProt database.