"Beke\u0161ov\u00E1, Sl\u00E1vka" . "17"^^ . "Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)" . "Komis, Georgios" . "[C05112ADBA9E]" . "\u0160amaj, Jozef" . . "Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)" . . . "10.1007/978-1-4939-0922-3_5" . "Clifton" . "http://link.springer.com/protocol/10.1007%2F978-1-4939-0922-3_5" . . "15310" . . "Protein phosphorylation is the most abundant and best studied protein posttranslational modification, dedicated to the regulation of protein function and subcellular localization as well as to protein-protein interactions. Identification and quantitation of the dynamic, conditional protein phosphorylation can be achieved by either metabolic labeling of the protein of interest with P-32-labeled ATP followed by autoradiographic analysis, the use of specific monoclonal or polyclonal antibodies against the phosphorylated protein species and finally by phosphoproteome delineation using mass spectrometry. Hereby we present a fourth alternative which relies on the enforced-affinity-based-electrophoretic separation of phosphorylated from non-phosphorylated protein species by standard SDS-PAGE systems co-polymerized with Phos-Tag (TM) and Mn2+ or Zn2+ cations. Phosphate groups of phosphorylated Ser, Thr, and Tyr residues form complexes with Mn2+ and Zn2+ cations with polyacrylamide immobilized PhosTag (TM). Following appropriate treatment of the gels, separated proteins can be quantitatively transferred to PVDF or nitrocellulose membranes and probed with common-not phosphorylation state specific-antibodies and delineate the occurrence of a certain phosphoprotein species against its non-phosphorylated counterpart." . . . "RIV/61989592:15310/14:33151290!RIV15-MSM-15310___" . . "Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)"@en . . . . "P(GAP501/11/1764), P(LO1204)" . "5"^^ . "immunoblotting; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Phos-TagTM; phosphorylation; Mitogen-activated protein kinase"@en . . . . "Vadovi\u010D, Pavol" . "Methods in Molecular Biology 1171" . "5"^^ . "Humana Press" . "266"^^ . . "Plant Map Kinases: Methods and Protocols" . "1801" . "Komis, Georgios" . . "Protein phosphorylation is the most abundant and best studied protein posttranslational modification, dedicated to the regulation of protein function and subcellular localization as well as to protein-protein interactions. Identification and quantitation of the dynamic, conditional protein phosphorylation can be achieved by either metabolic labeling of the protein of interest with P-32-labeled ATP followed by autoradiographic analysis, the use of specific monoclonal or polyclonal antibodies against the phosphorylated protein species and finally by phosphoproteome delineation using mass spectrometry. Hereby we present a fourth alternative which relies on the enforced-affinity-based-electrophoretic separation of phosphorylated from non-phosphorylated protein species by standard SDS-PAGE systems co-polymerized with Phos-Tag (TM) and Mn2+ or Zn2+ cations. Phosphate groups of phosphorylated Ser, Thr, and Tyr residues form complexes with Mn2+ and Zn2+ cations with polyacrylamide immobilized PhosTag (TM). Following appropriate treatment of the gels, separated proteins can be quantitatively transferred to PVDF or nitrocellulose membranes and probed with common-not phosphorylation state specific-antibodies and delineate the occurrence of a certain phosphoprotein species against its non-phosphorylated counterpart."@en . "978-1-4939-0921-6" . . . "RIV/61989592:15310/14:33151290" . . . . "Tak\u00E1\u010D, Tom\u00E1\u0161" . "Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)"@en .