User:Steven Moss/Notebook/AU CHEM-570/2012/02/21

Spring 2012 Biomaterials Design Lab

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Objective Purification and concentration of GFP Proceduree Purification of GFP with FPLC FPLC was run with a 5mL Ni column using the previously made binding and elution buffers 25mL of binding buffer (5X column volume) were run over the column 25-50 mL of GFP solution were then run on the column so GFP could bind to Ni 25 more mL of binding buffer were run on column 25 mL of elution buffer were run over column and fraction collection was started Process was repeated from beginning with 25 mL of binidng buffer over column until all GFP was collected. Concentration of GFP All fractions from FPLC that contained GFP were combined and concentrated including 3 washings with the binding buffer Data Notes All fractions containing GFP appeared green so no additional testing was done Started by doing 50 mL of GFP solution per column elution and moved towards doing lower volumes closer to 25 mL.