title: 2-D differential membrane proteome analysis of scarce protein samples.
abstract: 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 mug protein was found to be sufficient to detect proteins in a widespread well-separated diagonal spot pattern.
authors.affiliation: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Helling
authors.forename: Edgar
authors.initials: E
authors.lastname: Schmitt
authors.forename: Cornelia
authors.initials: C
authors.lastname: Joppich
authors.forename: Thomas
authors.initials: T
authors.lastname: Schulenborg
authors.forename: Stefan
authors.initials: S
authors.lastname: Müllner
authors.forename: Stephanie
authors.initials: S
authors.lastname: Felske-Müller
authors.forename: Thomas
authors.initials: T
authors.lastname: Wiebringhaus
authors.forename: Gabriele
authors.initials: G
authors.lastname: Becker
authors.forename: Gudrun
authors.initials: G
authors.lastname: Linsenmann
authors.forename: Barbara
authors.initials: B
authors.lastname: Sitek
authors.forename: Petra
authors.initials: P
authors.lastname: Lutter
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Meyer
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D020543:Proteome; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005456:Fluorescent Dyes; D008565:Membrane Proteins; D020543:Proteome