Method and kit for quantitative analysis of protein
US7354996B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jan 4, 2005 |
| Grant date | Apr 8, 2008 |
| Priority date | — |
| Expiry date | Nov 12, 2025 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/6851
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention provides a method for global quantitative analysis of protein that is effectively applied also for unpurified samples such as biological samples, and achieves better detection sensitivity and quantitativeness than the conventional NBS method. A method for global quantitative analysis of protein comprising: preparing two states of protein samples, a Protein sample I for analysis and a control Protein sample II; solubilizing the Protein sample I and II by urea or guanidine hydrochloride; subjecting the solubilized Protein sample I and II to modification using 2-nitro[13C6]benzenesulfenyl chloride and 2-nitro[12C6]benzenesulfenyl chloride; mixing and desalting the modified Protein sample I and II; resolubilizing by urea or guanidine hydrochloride; reducing and alkylating; subjecting to trypsin digestion in the presence of urea or guanidine hydrochloride; separating the obtained peptide mixture using a media having a phenyl group; and subjecting the enriched modified peptide fragments to mass spectrometry preferably using 3CHCA, 3H4NBA or mixture of 3H4NBA and 4CHCA as a matrix.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.