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Polyacrylamide lamination enables mass spectrometry compatible staining and in‐gel digestion of proteins separated by agarose IEF
Author(s) -
Hellman Jukka
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700299
Subject(s) - agarose , chromatography , polyacrylamide , silver stain , staining , chemistry , mass spectrometry , microbiology and biotechnology , biology , polymer chemistry , genetics
Abstract Agarose IEF enables the separation of large proteins and protein complexes. A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to separate the proteins, followed by layering the agarose with polyacrylamide. The formed laminate gels were seamless and durable and they were readily detached from the polyester. The gels were amenable to MS compatible staining. The sensitivity obtained with the acidic silver staining method was 20–50 ng/band of myoglobin. Laminated agarose was a suitable matrix for in‐gel digestion based generation of tryptic peptides for MALDI‐MS.