User:Susan Schultz/Notebook/Experimental Biological Chemistry/2011/09/20

Protein Extraction

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Objective Extract Maltose Binding Protein from ER2566 E.coli with the pMAL-pIII plasmid. Description See in pMal protein fusion and purification system manual. <http://www.neb.com/nebecomm/ManualFiles/manualE8000.pdf> The frozen cells in column buffer from the growth cultures prepared on 9/14/11 were thawed in a water bath. They were then sonicated in 15 second intervals for a total of 90 seconds, and were kept in ice between sonications. The mixtures were then centifuges for 30 minutes at 9000g. The supernatant, containing the protein was removed and purified using a column. Amylose Resin was poured into a 15mL column and it was washed with 120mL of column buffer. This column prep was run overnight. Data Add data and results here... Notes This area is for any observations or conclusions that you would like to note. Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.