Phos-tag SDS-PAGE analysis of <i>in vivo</i> phosphorylation of microtubule-associated protein tau and its protein kinase GSK3β

Phosphorylation is a posttranslational modification of proteins that regulates many biological functions, such as development, morphogenesis, gene expression, metabolism and neuronal activity. Dysregulation phosphorylation causes diseases including cancer Alzheimer’s disease. Then, lot research has been performed to reveal the molecular mechanism phosphorylation; identification site(s), protein kinases phosphatases, their regulation. Now, in vivo sites can be identified using mass spectrometry analyzed by immunoblotting with phospho-specific antibodies. However, it not easy grasp total profile target protein, when phosphorylated at multiple sites, quantification site-specific still challenging. Phos-tag SDS-PAGE phospho-affinity SDS-PAGE, which overcomes some above problems. Phosphorylated are delayed during electrophoresis separated from non-phosphorylated counterpart depending on number phosphorylation. We have employed this technique see proteins. Here, we introduce our results tau kinase GSK3β show how useful analysis