Dye ligand chromatography and two-dimensional electrophoresis of complex protein extracts from mouse tissue.
A complex protein fraction of mouse brain was subjected to dye ligand chromatography with various dye ligands. The proteins that were bound by the dye-gel matrix and also the non-binding proteins were separated by high-resolution two-dimensional electrophoresis. The protein patterns obtained were compared. The results show that a large number of different protein species bind to dye ligands and do not occur in the eluate. Red A was the most efficient dye in isolating an individual protein class from a complex tissue extract. Moreover, we found that many of the binding proteins did not cross-react among different types of dye ligands. Orange A and Blue B were the most unrelated dyes among those compared. Our investigation shows that dye ligand chromatography can be used as a means (among others employed previously) of fractionating and classifying the enormous number of different protein species in a mammalian tissue when combined with high-resolution two-dimensional electrophoresis.
Jungblut
P
P
Institut für Humangenetik, Freie Universität Berlin, F.R.G.
Klose
J
J
D000818:Animals; D001921:Brain / Q000378:metabolism; D002853:Chromatography, Liquid; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D051379:Mice; D011506:Proteins / Q000032:analysis*
D004396:Coloring Agents; D008024:Ligands; D011506:Proteins