FAQ: Will extended binding increase the yield of the target protein in the eluate?

In some cases, extending the binding step can result in significantly increased binding of the target protein.   Cap the column and seal the bottom using the plug provided. Mix end-over-end at 4°C for desired time (typically 5-15 minutes but can be extended overnight if necessary).  However, keep in mind that prolonged mixing may also result in increased non-specific protein binding.