User:Tamra L. Fisher/Notebook/Experimental Biological Chem/2012/03/28

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Objective

Continue the purification of the MBP-intein.

Description

  1. Analyze protein gel from yesterday to see where protein is present. (the protein MBP- Mxe GyrA intein -CBD fusion should be about 71kDa.
  2. Combine fractions 8-13, 30-35, 51-56, 72-77, and 92-97 in the concentrator.
  3. Concentrate this down to about 20mL.
  4. Add about 100mL of 50mM Tris (pH~9) to the concentrator to wash the excess salt out.
  5. Concentrate down to a final volume of about 10mL.
  6. Filter the protein and store at 4°C.




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