Electrophoretic analysis of phospho and non-phospho protein mixtures extracted from zebrafish whole brain samples by immobilized metal affinity electrophoresis

Phosphoproteins are principle cellular protein components with diverse regulatory functions and phosphorylation is the most frequent post-translational modifications of proteins. Immobilized metal affinity electrophoresis (IMAEP) is a recently developed technique by which the phosphoprotein components of the cellular samples could be captured. We have made use of this new methodology to capture the whole phosphoproteins of zebrafish brain. Since the elution and resolution of captured phosphoproteins by this new methodology are not yet quite developed, we have tried to make this new methodology more efficient in (1) capturing phosphoproteins from biological samples and (2) elution and resolution of captured phosphoproteins. In this project, we first examined the captured phosphoproteins from zebrafish whole brain samples, as a mixture of phosphoproteins and non-phosphoproteins, examined and resolved the captured phosphoproteins by electrophoresis, and finally eluted them successfully from the gel. In this work, we provided an efficient methodology for the elution of captured phosphoproteins from the gel which is an important development in IMAEP in the analysis of phosphoprotein component of cellular samples and showed the possibility of elution of the captured phosphoproteins. The developed methodology will potentially have wide applications in profiling phosphoproteins from biological samples like zebrafish brain and also in studies about signal transduction systems.