Use of high-performance size-exclusion chromatography to measure protein molecular weight and hydrodynamic radius. An investigation of the properties of the TSK 3000 SW column.
We have conducted a study of the TSK 3000 SW high-performance size-exclusion column to define under what conditions proteins would migrate most consistently with their known hydrodynamic properties. Our findings include the following: 1) the residual negative charge of the column does cause charge-exclusion or charge-retention effects at low ionic strengths; with elution in deionized water several anionic proteins elute approximately in the void volume; 2) at mu greater than or equal to 0.5, protein migration is not only independent of ionic strength, but consistent with protein molecular weight and hydrodynamic volume; 3) small hydrophobic peptides are retarded by the column; and 4) very asymmetric proteins and other hydrodynamic particles are likely to be retarded by an "end-on insertion" mechanisms.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel / Q000295:instrumentation; D008970:Molecular Weight; D011487:Protein Conformation; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis*; D012967:Sodium Dodecyl Sulfate
D000596:Amino Acids; D011506:Proteins; D012967:Sodium Dodecyl Sulfate