Further purify MBP with amylose column
- Create column buffer with 20 mM tris, 200 mM NaCl, and 1 mM EDTA (1.0 L made and filtered)
- Create elution buffer with column buffer + 10 mM Maltose (only need about 100 mL)
Regeneration of Amylose Resin Column
- Run 90mL of distilled water over the column at 4mL/min.
- Run 90mL of 0.1% SDS over the column at 4mL/min.
- Run 30mL of distilled water over the column at 4mL/min.
- Run 150mL of column buffer
- The filtered fractions showing protein (~10mL) were pumped over the regenerated amylose resin column at 1.0mL/min.
- Wash the column with 100mL of column buffer at 1.0mL/min.
- Elute the protein from the column with 100mL elution buffer at 1.0mL/min.