Gel Filtration Chromatography (Size Exclusion Chromatography) of Proteins

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.