Comparison between binding analyses performed by equilibrium dialysis and partitioning in aqueous two-phase systems exemplified by the binding of Cibacron Blue to serum albumin.
Partitioning in two-phase systems was used as a separation method in binding analyses and compared with equilibrium dialysis. A phase system was chosen to give partitioning conditions with the reactants in different phases. The interaction between serum albumin and Cibacron Blue F3G-A was used as a model system. The protein partitioned to the bottom phase and the ligand to the top phase in a phase system consisting of poly(ethylene glycol) and dextran. When both albumin and Cibacron Blue were present in the phase system, the degree of binding was observed as a translocation of ligand from the top to the bottom phase. An evaluation method that compensates for the amount of free reactant in the phase with the binding protein is presented.
D003956:Dialysis; D006801:Humans; D008956:Models, Chemical; D011485:Protein Binding; D012709:Serum Albumin; D012995:Solubility; D014227:Triazines
D012709:Serum Albumin; D014227:Triazines; C012842:Cibacron Blue F 3GA