Bio-Rad Profinity IMAC Resins - Manual (Page 19)

3. Sonicate the lysate (on ice) 4 times at 1 min intervals.
4. Centrifuge the lysate at 12,000 x g for 20 min at
4°C.
5. Remove the supernatant, and filter through a
0.2 µM filter immediately before applying to the
cartridge.
Denatured Lysates
1. Harvest cell pellet by centrifugation at 8,000 x g for
10 min at 4°C.
2. Determine weight of pellet and resuspend in
10 volumes denaturing lysis/wash buffer 1 (200 ml of
culture typically yields 0.8 g of paste, and results in
8 ml of lysate).
3. Sonicate the lysate 4 times at 1 min intervals.
4. Centrifuge the lysate at 12,000 x g for 20 min at
4°C.
5. Remove the supernatant, and filter through a
0.2 µM filter immediately before applying to the
cartridge.
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