High-performance molecular sieve chromatography of proteins on agarose columns: the relation between concentration and porosity of the gel.
Curves showing the relation between log (molecular weight) and distribution coefficient are presented for proteins subjected to molecular sieve chromatography on crosslinked and non-crosslinked agarose gels of different concentrations. These curves, which facilitate selection of the gel concentration that gives optimal resolution in any particular separation problem, show that the exclusion limit of 5, 9, 12, and 20% agarose gels correspond to protein with molecular weights above 1,000,000, 600,000, 450,000, and 280,000, respectively. Plate numbers have been determined for columns of 20% agarose at different flow rates and bead sizes. Separations of model proteins by high-performance molecular sieve chromatography on agarose beads are shown.
D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008970:Molecular Weight; D010316:Particle Size; D010751:Phosphopyruvate Hydratase / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012709:Serum Albumin / Q000032:analysis
D011506:Proteins; D012709:Serum Albumin; D010751:Phosphopyruvate Hydratase