Shotgun proteomics analysis of PLBD2
Both human and CHO PLBD2 samples were subjected to shotgun proteomics analysis. 10 µg of PLBD2 was dried with Speedvac, then re-constituted with 20 µl of denature/reduction buffer containing 8M urea and 10 mM DTT. The proteins were denatured and reduced at 37°C for 30 min, and then incubated with 6 µl of 50 mg/ml iodoacetamide for 30 min in dark. Alkylated proteins were digested overnight with 50 µl 0.01 µg/µL trypsin at 37 °C. The peptide mixture was acidified by 5 µL of 10% TFA. The sample was injected 10 µL for LC-MS/MS analysis.