Naslov
Use of selective extraction and fast chromatographic separation combined with electrophoretic methods for mapping of membrane proteins

Autori
Josi, Đuro ; Brown, Mari Kino ; Huang, Feilei ; Callanan, Helen ; Ručević, Marijana ; Nicoletti, Alison ; Clifton James ; Hixson, Douglas C.

Izvornik
Electrophoresis (0173-0835) 26 (2005), 14; 2809-2822

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
convective interaction media monoliths; plasma membranes; protein mapping

Sažetak
A model system for selective solubilization and fast separation of proteins from the rat liver membrane fraction and purified rat liver plasma membranes for their further proteomic analysis is presented. For selective solubilization, high-pH solutions and a concentrated urea solution, combined with different detergents, are used. After extraction, proteins are separated by anion-exchange chromatography or a combination of anion- and cation-exchange chromatography with convective interaction monolithic supports. This separation method enables fast and effective prefractionation of membrane proteins based on their hydrophobicity and charge prior to one-dimensional (1-D) and 2-D electrophoresis and mass spectrometry. By use of this sample preparation method, the less-abundant proteins can be detected and identified.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

Napomena
Rad ostvaren u suradnji s: Proteomics Core, COBRE Center for Cancer Research Development, Rhode Island Hospital, The CORO Center, Providence, RI, USA i Division of Medical Oncology, Department of Medicine, Rhode Island Hospital, Providence, RI, USA

POVEZANOST RADA