Method optimization for proteomic analysis of soybean leaf: Improvements in identification of new and low-abundance proteins.

ABSTRACT: The most critical step in any proteomic study is protein extraction and sample preparation. Better solubilization increases the separation and resolution of gels, allowing identification of a higher number of proteins and more accurate quantitation of differences in gene expression. Despite the existence of published results for the optimization of proteomic analyses of soybean seeds, no comparable data are available for proteomic studies of soybean leaf tissue. In this work we have tested the effects of modification of a TCA-acetone method on the resolution of 2-DE gels of leaves and roots of soybean. Better focusing was obtained when both mercaptoethanol and dithiothreitol were used in the extraction buffer simultaneously. Increasing the number of washes of TCA precipitated protein with

SUBMITTER: Mesquita RO 

PROVIDER: S-EPMC3392888 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature