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FAQ: My purification failed and/or why is my yield lower than expected?

Multiple reasons may account for purifications resulting in low yield. It is important to first optimize the expression level of the soluble target protein by altering induction time/ temperature and level of inducer. It is possible that the protein has formed inclusion bodies or the tag has been sequestered and therefore is unable to bind to the beads. In this case, the purification should be performed under denaturing conditions. Insufficient cell lysis and extraction will also lead to reduced yield; it is important to ensure your lysis procedure is sufficient. In some cases, extending the binding time may improve protein binding.