Bio-Rad Profinity IMAC Resins - Manual (Page 15)

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6. Adjust the adaptor to sit directly on top of resin bed.
Open the column outlet and pump distilled buffer through the column at a
packing flow rate of ~400–600 cm/hr for 5 to 10 min or at the maximum
pressure allowed by the column hardware and resin. While packing at high
flow rates, the resin bed will be compressed down. Mark this
compression level with a pen.
Stop the flow. At this point, the resin bed height might readjust and rise up. If
this happens, adjust the flow adaptor to compress the bed an additional
0.1–0.5 cm down past the level marked with the pen.
7. Reconnect the pump and equilibrate.
Pass eluent (distilled water) through the column at the packing flow rate.
During equilibration, the bed might be compressed down even further. After a
constant bed height is reached, mark the compression level at this flow rate.
Again, adjust the adaptor to further compress the bed an additional
0.1–0.5 cm past the level marked.
8. Proceed with Immobilizing Metal Ions procedure (Section 6) if using
uncharged Profinity IMAC resin or if recharging Profinity IMAC
Ni-charged resin.
Note: Chromatographic steps during purification should not be run greater
than 75% of the packing flow rate.
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