User:Steven Moss/Notebook/AU CHEM-570/2012/04/03: Difference between revisions
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==Objective== | ==Objective== | ||
Further purify MBP with amylose column | |||
== | ==Procedure== | ||
# Create column buffer with 20 mM tris, 200 mM NaCl, and 1 mM EDTA | |||
# 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. | |||
==Data== | ==Data== |
Revision as of 11:50, 3 April 2012
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ObjectiveFurther purify MBP with amylose column Procedure
Regeneration of Amylose Resin Column
Purifying MBP
DataNotes |