Bio-Rad Profinity IMAC Resins - Manual (Page 18)

Section 7
Preparation of E. coli
Lysates
For E. coli cultures expressing medium to high levels of
His-tagged proteins, (
≥10% of total protein), 200 ml of
culture will yield sufficient material for a 1 ml cartridge
purification, and 1,000 ml of culture will yield sufficient
material for a 5 ml cartridge purification run. For cultures
expressing protein at low levels (
≤10% of total protein),
the culture volumes will need to be determined
empirically for each protein.
Native 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 native lysis/wash buffer 1 (200 ml of
culture typically yields 0.8 g of paste, and results in
8 ml of lysate).
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