Establishment of two-dimensional gel electrophoresis system for myofibrillar proteins of silver carp (Hypophthalmichthys molitrix)

A two-dimensional gel electrophoresis (2-DE) system for the separation of myofibrillar proteins from silver carp was established and optimized in this study. The key factors of isoelectric focusing (IEF) and the pH gradient of immobilized pH gradient (IPG) strip were selected as the main factors affecting 2-DE in this paper. We loaded 120 μg of protein onto the 17 cm pH 4-7 IPG strip, and the IEF program III was as follows: 50 V for 15 h, three-stage desalting involving 250 V for 2 h, 1 000 V for 2.5 h and 4 000 V for 3 h, 8 000 V for 2 h and 10 000 V for 1.5 h, and 10 000 V for 80 000 V·h and 500 V held for 10 h. The gel concentration was 12%. The gel was stained with sliver nitrate, scanned and analyzed using PDQuest software. A high-resolution 2-DE map for myofibrillar proteins from silver carp was obtained finally. © 2017, China Food Publishing Company. All right reserved.