Optimization of Two-dimensional Polyaneylamide Gel Electrophoresis for Proteome of Kandelia candel Roots

A two-dimensional polyarelamide gel electrophoresis (2D-PAGE) system was established for proteomic analysis of Kandelia candel roots. Protein extraction, dissolution, loading quantity, IEF and SDS-PAGE parameters in the two-dimensional electrophoresis (2-DE) were optimized. The results showed that phenol method was suitable for the root protein extraction, the protein isoelectric points mainly distributed between pH 4 and 7. The lysis buffer with 2 mmol稬-1 thiourea improved dissolution of protein effectively and the resolution of spectrum for the dissolved protein was increased with 40 mmol稬-1 DTT and 1.5 mg loading amount. The 1089 protein spots were stained with Colloidal coomassie blue and detected by mass spectrometry.