Bio-Rad Profinity IMAC Resins - Manual (Page 21)

17
Note: Some portion of the target protein will remain insoluble and will localize
in the pellet. To properly release the His-tagged protein from the pellet, it will
need to be subjected to denaturing conditions. See Section 8, Preparation of
Clarified E. coli Lysate Using Denaturing Conditions.
7. Analyze the concentration of the target protein using a small amount of
supernatant.
Use SDS-PAGE to get a rough determination of the protein’s concentration.
This may help determine the appropriate size of the IMAC column needed.
8. Continue with purification protocols using nondenaturing conditions.
Proceed with protocols using nondenaturing conditions.
10001677B.qxd 1/28/2005 12:42 PM Page 20