FAQ: I’ve prepared my lysate with too much buffer and the target is very dilute, can I apply more lysate by repeating the load steps?

Yes, if the lysate volume is larger than 500 μl, multiple applications to the same column can be performed.  Optimal target binding to the resin is related to the concentration of the target, so in some cases, where significant dilution has occurred, it may be necessary to prepare a more concentrated lysate for optimum binding of the target protein.