Determination of Triton X-100 binding to membrane proteins by polyacrylamide gel electrophoresis.
The molecular weight of proteins in protein-detergent complexes can be determined from ultracentrifugation experiments if the amount of bound detergent is known. A new sensitive method to measure the binding of the nonionic detergent Triton X-100 to proteins has been developed. For the membrane proteins studied, less than 50 mug of protein was required to achieve an accuracy of 10% in the determination of the detergent-protein weight ratio. The proteins were equilibrated with the detergent by electrophoresis into polyacrylamide gels containing radioactively labelled Triton X-100. The gels were then sliced and the amount of bound detergent calculated from the increase in radio-activity in the slices containing the protein zone. The amounts of protein were determined by amino acid analysis of identical protein zones cut from gels running parallel.
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008565:Membrane Proteins; D008970:Molecular Weight; D011092:Polyethylene Glycols; D011485:Protein Binding; D011506:Proteins; D000644:Quaternary Ammonium Compounds
D008565:Membrane Proteins; D011506:Proteins; D000644:Quaternary Ammonium Compounds; D011092:Polyethylene Glycols