title: Time-based analysis of silver-stained proteins in acrylamide gels.
abstract: Silver staining of proteins after PAGE often remains the method of choice in many laboratories. Nevertheless, it is known that quantification of protein levels is keenly restricted to a small range of protein concentrations leading to an over- or underestimation of protein amounts. To overcome this, a time-based analysis method was developed to avoid the saturation effect of the silver-staining reaction, thus resulting in an improved dynamic range of the gel image produced and therefore better quantification of proteins. Instead of the well-known end-point image analysis, gray intensities of time series images of a developing gel are determined and times until a threshold gray value is reached are calculated. These times are used to calculate a new grayscale image which can be analyzed using commercial image processing software.
authors.affiliation: Department of Cardiovascular Medicine, Institute of Physiology, University of Greifswald, Karlsburg, Germany. bertram.becher@uni-greifswald.de
authors.forename: Bertram
authors.initials: B
authors.lastname: Becher
authors.forename: Ann-Kathrin
authors.initials: AK
authors.lastname: Knöfel
authors.forename: Jörg
authors.initials: J
authors.lastname: Peters
mesh_terms: D000818:Animals; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods* / Q000706:statistics & numerical data; D007091:Image Processing, Computer-Assisted; D011506:Proteins / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis; D012834:Silver; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D011506:Proteins; D012710:Serum Albumin, Bovine; D012834:Silver