The Sample Preparation and the Improvement Method of Two-Dimensional Electrophoresis of Proteins from Antiaris toxicaria (Moraceae) Seed

Abstract

We have established the extraction method of the total proteins of Antiaris toxicaria seed and the appropriate conditions for the two-dimensional eiectrophoresis(2-DE) of Antiaris toxicaria seed proteome. Through integrating and optimizing various conditions,we also established the 2-DE system including the sample preparation method with Tris-HCL based on TCA-actone,and the first-dimensional electrophoresis with immobilized pH gradients isoelectric focusing and the second dimensional electrophoresis with vertical electrophoresis of SDS-PAGE. We have gained a good 2-DE map in the analysis of sample preparation、sample resolution、isoelectric focusing(IEF) 、SDS-PAGE and gel staining. In order to explore the extraction method of proteome and technique of 2-DE,we compared the extraction method between the TCA-actone and the Tris-HCl,and improved the key steps of 2-DE. The Tris-HCl extraction was the most appropriate for Antiaris toxicaria seed proteome analysis because of the highest resolution and more informative 2-DE map. Those were the solid basis for the later research of studying on the discrepancy expression proteomics of Antiaris toxicaria Seed.