title: On the preparation and binding characteristics of Cibacron blue-glycerolpropyl-glass.
abstract: The dye Cibacron blue, which has a binding affinity for the 'dinucleotide fold' in enzymes using dinucleotide coenzymes, was covalently immobilized on glycerolpropyl-glass. Derivatives of controlled-pore glass beads (1489 A pore diam.; 80/120 mesh) were formed to give a glycerol surface phase by treatment with gamma-glycidoxypropyltrimethoxysilane. The method of Taylor and Swaisgood (Biotechnol. Bioeng. 23 (1981) 1349-1363) was used to determine simultaneously the partition coefficient and the dissociation constant for the affinity ligand using heart lactate dehydrogenase. Nonspecific partitioning of the enzyme in the glycerolpropyl-matrix did not occur, thus K = 1. The dissociation constant for binding of the enzyme to the immobilized ligand was 2.53 microM, which is an excellent agreement with that determined for interaction with the soluble ligand.
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D005898:Glass; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology; D014227:Triazines
chemical_list: D014227:Triazines; C012842:Cibacron Blue F 3GA; D007770:L-Lactate Dehydrogenase