In-gel tagging and in-gel digestion for phosphoproteins analysis and phosphorylation site identification
US7629744B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 8, 2004 |
| Grant date | Dec 8, 2009 |
| Priority date | — |
| Expiry date | Mar 29, 2025 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/6848
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention relates to a method for phosphorylation site-specific labeling of phosphoproteome with a site-specific tagging reagent and analyzing of the resulting labeled one, more especially, a method for in-situ tagging of phosphorylation sites of phosphoproteins retained in polymeric gel with a nucleophilic tagging reagent. It also relates a method for generating new proteolytic cleavable sites at formerly phosphorylation sites by a proper choice of a nucleophilic tagging reagent. It also relates to a method for phosphopeptides analysis and phosphorylation site identification by in-gel digestion of the previously in-gel tagged proteins and subsequent mass analysis of the resulting peptides. The invention provides in-gel chemical tagging method for phosphoaminoacid residue of phosphoproteins retained in polymeric gel matrix. Phosphoprotein can be immobilized into gel matrix by a variety of methods such as gel electrophoresis. The immobilized phosphoproteins are retained in gel matrix during tagging reaction to phosphorylated aminoacid residue of phosphoproteins, and the resulting tagged proteins are also retained in gel matrix till following purification steps like washi…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.