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

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.