title: Gel filtration chromatography (size exclusion chromatography) of proteins.
abstract: The protocol described here allows the student to construct a standard curve for a gel filtration column with a separation range of 5-250 kD. The size (hydrodynamic radius) of a protein species stable in a buffer containing Tris-HCl, NaCl, and DTT is determined using this column. Modifications may be made to the buffer to accommodate the protein of interest and the separation range of the column.
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Krisna C
authors.initials: KC
authors.lastname: Duong-Ly
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: gabelli@jhmi.edu.
authors.forename: Sandra B
authors.initials: SB
authors.lastname: Gabelli
mesh_terms: D002021:Buffers; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods*; D011506:Proteins / Q000302:isolation & purification*; D014325:Tromethamine / Q000737:chemistry
chemical_list: D002021:Buffers; D011506:Proteins; D014325:Tromethamine
keywords: Column volume; Gel filtration chromatography; Gel filtration column; Size exclusion chromatography; Void volume