title: 2D-ToGo workflow: increasing feasibility and reproducibility of 2-dimensional gel electrophoresis.
abstract: Two-dimensional gel electrophoresis (2-DE) is one of the most powerful methods for studying global protein profiles. However, due to the multiple manual steps involved in gel based processing it is challenging to achieve the necessary overall reproducibility for a reliable comparative analysis, especially between different laboratories. To improve the 2-DE technique for quantitative analyses we have set up a robust 2-DE workflow, called 2D-ToGo, which utilizes latest innovations concerning instrumentation, consumables and protocols. Quantitative data analyses indicate the high reproducibility between replicate gels processed at a single site (intra-laboratory variation: CV 20%). The data-sets of the inter-laboratory comparison revealed similar results displaying a variation of CV 23%. The technical improvements given by our 2-DE workflow have a positive impact on process robustness and most importantly, reproducibility. Accordingly, many of the well-known challenges for resolving and quantitating up to thousands of different protein components in a given biological sample are minimized.
authors.affiliation: Bio-Rad Laboratories GmbH, Munich, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Posch
authors.forename: Thomas
authors.initials: T
authors.lastname: Franz
authors.forename: Sonja
authors.initials: S
authors.lastname: Hartwig
authors.forename: Birgit
authors.initials: B
authors.lastname: Knebel
authors.forename: Hadi
authors.initials: H
authors.lastname: Al-Hasani
authors.forename: Waltraud
authors.initials: W
authors.lastname: Passlack
authors.forename: Nancy
authors.initials: N
authors.lastname: Kunz
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Hinze
authors.forename: Xinping
authors.initials: X
authors.lastname: Li
authors.forename: Jorg
authors.initials: J
authors.lastname: Kotzka
authors.forename: Stefan
authors.initials: S
authors.lastname: Lehr
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods / Q000592:standards*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000032:analysis*; D005612:Freeze Drying; D007091:Image Processing, Computer-Assisted; D007525:Isoelectric Focusing; D015588:Observer Variation; D015203:Reproducibility of Results; D057188:Workflow
chemical_list: D029968:Escherichia coli Proteins