An electrophoretic study on interactions of albumins of different species with immobilized Cibacron Blue F3G A.
Albumins of different species, varying in electrophoretic mobility, were compared in their interaction with the dye Cibacron Blue F3G A. Immobilized by coupling to a high molecular weight dextran ("blue dextran"), the dye was used as a ligand in affinity electrophoresis in different setups. One-dimensional electrophoresis with blue dextran entrapped in an intermediate gel and two-dimensional applications with transverse gradients (affinity titration curves, zonal electrophoresis in linear ligand gradients) were performed. Compared to the human homologue, animal albumins albumins display more complex patterns and interaction profiles, depending on pH and ionic strength of the buffers. Results may differ considerably from those obtained by affinity chromatography, illustrating the additional screening potential of the electrophoretic methods. Comparison of different samples under the influence of ligand competition, reducing conditions, or denaturing agents gives supplementary information on conformational behavior of the proteins.
Miller
I
I
Institut für Medizinische Chemie, Veterinärmedizinische Universität Wien, Austria. ingrid.miller@vuwien.ac.at
Gemeiner
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M
D000418:Albumins / Q000737:chemistry*; D000818:Animals; D002021:Buffers; D002415:Cats; D002417:Cattle; D002846:Chromatography, Affinity; D004396:Coloring Agents; D004285:Dogs; D004586:Electrophoresis / Q000379:methods*; D006736:Horses; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D013045:Species Specificity; D013552:Swine; D014227:Triazines / Q000737:chemistry*
D000418:Albumins; D002021:Buffers; D004396:Coloring Agents; D014227:Triazines; C012842:Cibacron Blue F 3GA