Addition of proteins to the cylindrical gel embedding medium for transverse molecular-weight markers in two-dimensional gel electrophoresis.
As an aid in the comparison of different complex mixtures of proteins resolved by two-dimensional electrophoresis, a simple method which results in the electrophoresis of molecular-weight standards as appropriately migrating, highly resolved bands extending across the entire second-dimension slab gel is described. The proteins to be used as markers are included in the molten agarose mixture used to affix the first-dimension cylindrical gel atop the second-dimension slab gel. As the proteins which are resolved in the first dimension migrate through the second-dimension slab gel, the marker proteins also migrate, experiencing the same electrophoretic conditions as the sample proteins in the immediate vicinity. If the same protein is resolved in the first dimension and also used as a marker, it electrophoreses in the second dimension as a spot intersected by a band traversing the entire gel. This sensitive method is applied to a comparison of soluble seed proteins of two cotton species, Gossypium hirsutum and G. arboreum, using G. hirsutum seed protein as the molecular-weight marker. Other applications are described.
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D003368:Gossypium; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010940:Plant Proteins / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012995:Solubility
D010940:Plant Proteins; D011506:Proteins