Bio-Rad Profinity IMAC Resins User Manual
Page 40
36
Remove remaining unbound proteins by centrifuging. Save flow-through and pool
with unbound fraction from step 6. Label “unbound 2”. The wash step can be
repeated if necessary.
Part 3: Eluting the His-Tagged Protein
10. Insert micro spin column into new, clean collection vessel.
11. Elute bound proteins with 5 column volumes of elution buffer.
Pipet up and down at least 5 times and incubate for 5 min.
12. Centrifuge at 1,000 x g for 1 min to recover His-tagged protein.
Analyze fractions from above steps by A
280
, SDS-PAGE, ELISA, etc.
10001677B.qxd 1/28/2005 12:42 PM Page 39
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