title: Western Blotting Using PVDF Membranes and Its Downstream Applications.
abstract: Western blotting using polyvinylidene difluoride (PVDF) membranes is one of the most popular techniques for detection and characterization of proteins. If this technique is combined with immunodetection, the behavior of a particular protein can be clarified. On the other hand, if it is combined with Edman sequencing, the primary structure of the protein can be determined. A protein sample is transferred from an SDS-polyacrylamide gel electrophoresis (PAGE) gel onto a PVDF membrane by electroblotting. The membrane carrying the protein is either used for immunodetection or protein sequencing. SDS-PAGE followed by Western blotting combined with immunodetection using antibodies can easily detect protein behavior in crude protein mixtures. Furthermore, two-dimensional PAGE followed by Western blotting and Edman sequencing allows effective sequence determination of crude protein mixtures that may not be easily purified by conventional column chromatography.
authors.affiliation: National Institute of Crop Science, NARO, Kannondai 2-1-18, Tsukuba, 305-8518, Japan, skomatsu@affrc.go.jp.
authors.forename: Setsuko
authors.initials: S
authors.lastname: Komatsu
mesh_terms: D000906:Antibodies / Q000737:chemistry; D015153:Blotting, Western / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006735:Horseradish Peroxidase / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D011145:Polyvinyls / Q000737:chemistry*; D011761:Pyrrolidonecarboxylic Acid / Q000737:chemistry; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry; D013912:Threonine / Q000737:chemistry; D005465:Fluorocarbon Polymers
chemical_list: D000906:Antibodies; D006735:Horseradish Peroxidase; D008567:Membranes, Artificial; D011145:Polyvinyls; D011761:Pyrrolidonecarboxylic Acid; D012694:Serine; D013912:Threonine; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers