Using differential solubilization and 2-D gel electrophoresis to visualize increased numbers of proteins in the human cortex and caudate nucleus and putamen.

ABSTRACT: The aim of this study was to determine if differential solubilization of human CNS proteins would increase the total number of proteins that could be visualized using 2-D gel electrophoresis. Hence, proteins were solubilized into Tris, CHAPS and SB3-10 before separation across a pH 4-7 IEF gradient and a 12-14% SDS polyacrylamide gel, which could be achieved with a run-to-run variation of 35% in spot intensity. Because Western blot analyses suggested proteins could be in more than one detergent fraction, we completed a conservative analyses of our 2-D gels assuming spots that appeared on multiple gels at the same molecular weight and pI were the same protein. These analyses show that we had visualized over 3000 unique protein spots across three 2-D gels generated from each sample of human

SUBMITTER: Dean B 

PROVIDER: S-EPMC3056166 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature