Gel-assisted Digestion [28] : Acrylamide/bisacrylamide (40%, 29:1), 10% APS, and TEMED were then applied to protein solution to polymerize as a gel directly in the eppendorf. The gel was cut into small pieces and washed several times with TEABC containing 50% ACN. The gel samples were further dehydrated with ACN and the completely dried by vacuum centrifugation. Proteolytic digestion was then performed with trypsin (protein: trypsin = 50:1, g/g) in 25 mM TEABC with incubation overnight at 37uC and peptides were extracted by 5% FA in 50% ACN, dried in a SpeedVac and stored in 230uC.