Celb2 Protein Chromatography Lab Report

In order to purify CelB2 protein bound to maltose-binding-protein, amylose affinity chromatography was performed. The amylose resin present in 20% ethanol was first diluted by adding 10 mL of 20 mM TrisHCl, pH 7.4, 0.2 M NaCl, 1 mM EDTA and centrifuged at 700 rpm for 5 minutes. After decanting the buffer, another 10 mL of TrisHCl, pH 7.4, 0.2 M NaCl, 1 mM EDTA was added to this resin solution and centrifuged at 700 rpm for another 5 minutes to further dilute the ethanol concentration in the resin. After the buffer was decanted again, the column was packed using the batch method, where the 17.9 mL of lysate solution containing the protein of interest was added to the resin. This solution was incubated for 1 hour, with shaking at 4ºC. The column