Antibody array for the detection of protein phosphorylation localization and application to dissect activities of different signaling pathways

摘要

This thesis Develop a new suitable nano-gold mediated silver antibody microarray technology. Application examples show that the antibody array efficently detects differences in protein and phospho-protein concentration independently of cell type. For activation and inhibition of signaling pathways, cytoplasmic and nuclear distribution are very important and can reflect differential functions, even when total phospho-proteins are not changed. Evidence for new and rapid common cytoplasmic signaling about ERK, JNK, STAT3, Akt and p70S6 in chosen functions of three treated cell lines. Setup new positive or negative co-localized relationships and interaction about p-p38 and p38, Smad1 and Smad2/3, ERK or p38 with Smad1, ERK or p38 with c-Myc, among Akt, ERK and p70S6, p70S6 with p38 and smad1,2,3 in three treated cell lines.