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Ultrafiltration for proteomic sample preparation
Author(s) -
Chernokalskaya Elena,
Gutierrez Sara,
Pitt Aldo M.,
Leonard Jack T.
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200405998
Subject(s) - ultrafiltration (renal) , chromatography , sample preparation , sample (material) , chemistry
Proteome analysis represents significant challenges to the existing sample preparation techniques. Traditional methods, such as two‐dimensional electrophoresis, typically separate high‐molecular‐weight proteins while discarding low‐molecular‐weight species. This approach is well justified considering the complexity of any proteome. However, it is desirable to extract the maximum amount of information from each sample to investigate the entire range of biomolecules. We have demonstrated that ultrafiltration not only improves two‐dimensional electrophoresis (2‐DE) resolution of the protein fraction but also yields the low‐molecular‐weight fraction amenable for further analysis by high‐resolution mass spectrometry. This approach was successfully adapted to the variety of biological samples including cell and tissue lysates and serum. Therefore, ultrafiltration offers an alternative sample preparation technique that enables more thorough analysis of a proteome.