title: Quantitative gel electrophoresis: new records in precision by elaborated staining and detection protocols.
abstract: Gel electrophoresis (GE) is a very common analytical technique for proteome research and protein analysis. Despite being developed decades ago, there is still a considerable need to improve its precision. Using the fluorescence of Colloidal Coomassie Blue -stained proteins in near-infrared (NIR), the major error source caused by the unpredictable background staining is strongly reduced. This result was generalized for various types of detectors. Since GE is a multi-step procedure, standardization of every single step is required. After detailed analysis of all steps, the staining and destaining were identified as the major source of the remaining variation. By employing standardized protocols, pooled percent relative standard deviations of 1.2-3.1% for band intensities were achieved for one-dimensional separations in repetitive experiments. The analysis of variance suggests that the same batch of staining solution should be used for gels of one experimental series to minimize day-to-day variation and to obtain high precision.
authors.affiliation: Institute of Pharmaceutical Chemistry, Technical University of Braunschweig, Braunschweig, Germany.
authors.forename: Xi
authors.initials: X
authors.lastname: Deng
authors.forename: Simone
authors.initials: S
authors.lastname: Schröder
authors.forename: Sabine
authors.initials: S
authors.lastname: Redweik
authors.forename: Hermann
authors.initials: H
authors.lastname: Wätzig
mesh_terms: D000704:Analysis of Variance; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000592:standards*; D005453:Fluorescence; D007091:Image Processing, Computer-Assisted; D011506:Proteins / Q000032:analysis; D012015:Reference Standards; D015203:Reproducibility of Results; D012394:Rosaniline Dyes / Q000737:chemistry; D013055:Spectrophotometry, Infrared; D013194:Staining and Labeling / Q000379:methods / Q000592:standards*; D013696:Temperature
chemical_list: D011506:Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue