FAQ: How can I determine if my target protein remains bound to the resin?

Remove an aliquot of the resin from the column and add 200 µL of SDS-PAGE loading buffer. Incubate at 70°C for 5 minutes to release any protein that remains on the resin following elution. Load and analyze proteins by SDS-PAGE. Most proteins, regardless of whether they bind to the nickel or to the agarose-bead itself, will be recovered by this procedure.