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.