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2‐D differential membrane proteome analysis of scarce protein samples
Author(s) -
Helling Stefan,
Schmitt Edgar,
Joppich Cornelia,
Schulenborg Thomas,
Müllner Stefan,
FelskeMüller Stephanie,
Wiebringhaus Thomas,
Becker Gabriele,
Linsenmann Gudrun,
Sitek Barbara,
Lutter Petra,
Meyer Helmut E.,
Marcus Katrin
Publication year - 2006
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.200600169
Subject(s) - proteome , membrane protein , chromatography , chemistry , membrane , proteomics , biochemistry , biology , gene
Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X‐114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D‐CTAB/SDS‐PAGE. For a differential analysis 3 μg protein was found to be sufficient to detect proteins in a widespread well‐separated diagonal spot pattern.