User:Steven Moss/Notebook/AU CHEM-570/2012/04/03

Spring 2012 Biomaterials Design Lab

Main project page Previous entry      Next entry

Objective Further purify MBP with amylose column Procedure 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 Purifying MBP 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.