title: A new procedure for fast soft staining of BN-PAGEs on photosynthetic complexes.
abstract: We report a fast and sensitive procedure for blue native PAGE staining, in which the conventional staining step with CBB is avoided. After running, a short exposure to a mix of polar protic solvents (ethanol and acetic acid) leads to a fast and selective removal of the dye from the migration front and a specific binding to the protein bands, while the rest undergo a selective and complete background removal, leading to an intense contrast. This single-step staining-destaining technique is useful in protein samples that bind colored cofactors such as photosystems, which can be selectively discerned by their characteristic green color. After the staining of such samples, the green color persists, while the other unpigmented protein complexes and the molecular standard remain CBB stained, creating a useful reference system for the assignment of the bands. The advantages and chemical basis of this staining procedure are discussed.
authors: Laboratory of Plant Physiology and Photobiology, Department of Life and Environmental Sciences, University of Cagliari, Cagliari, Italy.|Department of Molecular Sensory Systems, Center of Advanced European Studies and Research (Caesar), Bonn, Germany.
authors: Domenica
authors: D
authors: Farci
authors: Department of Core Facility Proteomics, Leibniz Institute on Aging - Fritz Lipmann Institute, Jena, Germany.
authors: Joanna
authors: J
authors: Kirkpatrick
authors: Laboratory of Plant Physiology and Photobiology, Department of Life and Environmental Sciences, University of Cagliari, Cagliari, Italy.
authors: Dario
authors: D
authors: Piano
mesh_terms: D063347:Native Polyacrylamide Gel Electrophoresis / Q000379:methods*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000032:analysis* / Q000737:chemistry*; D012394:Rosaniline Dyes / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*; D020524:Thylakoids / Q000737:chemistry; D014026:Nicotiana / Q000737:chemistry / Q000166:cytology
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue
keywords: BN-PAGE; Coomassie staining; Photosynthetic complexes; Pigmented proteins; Thylakoids