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| rdf:type
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| rdfs:seeAlso
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| Description
| - 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.
- 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)
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| Title
| - Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)
- Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM) (en)
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| skos:prefLabel
| - Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM)
- Affinity-Based SDS PAGE Identification of Phosphorylated Arabidopsis MAPKs and Substrates by Acrylamide Pendant Phos-Tag (TM) (en)
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| skos:notation
| - RIV/61989592:15310/14:33151290!RIV15-MSM-15310___
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| http://linked.open...avai/riv/aktivita
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| http://linked.open...avai/riv/aktivity
| - P(GAP501/11/1764), P(LO1204)
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| http://linked.open...vai/riv/dodaniDat
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| http://linked.open...aciTvurceVysledku
| |
| http://linked.open.../riv/druhVysledku
| |
| http://linked.open...iv/duvernostUdaju
| |
| http://linked.open...titaPredkladatele
| |
| http://linked.open...dnocenehoVysledku
| |
| http://linked.open...ai/riv/idVysledku
| - RIV/61989592:15310/14:33151290
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| http://linked.open...riv/jazykVysledku
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| http://linked.open.../riv/klicovaSlova
| - immunoblotting; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Phos-TagTM; phosphorylation; Mitogen-activated protein kinase (en)
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| http://linked.open.../riv/klicoveSlovo
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| http://linked.open...ontrolniKodProRIV
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| http://linked.open...i/riv/mistoVydani
| |
| http://linked.open...vEdiceCisloSvazku
| - Methods in Molecular Biology 1171
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| http://linked.open...i/riv/nazevZdroje
| - Plant Map Kinases: Methods and Protocols
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| http://linked.open...in/vavai/riv/obor
| |
| http://linked.open...ichTvurcuVysledku
| |
| http://linked.open...v/pocetStranKnihy
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| http://linked.open...cetTvurcuVysledku
| |
| http://linked.open...vavai/riv/projekt
| |
| http://linked.open...UplatneniVysledku
| |
| http://linked.open...iv/tvurceVysledku
| - Takáč, Tomáš
- Šamaj, Jozef
- Bekešová, Slávka
- Komis, Georgios
- Vadovič, Pavol
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| number of pages
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| http://bibframe.org/vocab/doi
| - 10.1007/978-1-4939-0922-3_5
|
| http://purl.org/ne...btex#hasPublisher
| |
| https://schema.org/isbn
| |
| http://localhost/t...ganizacniJednotka
| |
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