Dear All ,
I was purifying the proteins, following ProBond™ Purification System protocol (denaturing conditions) and noticed that Ni-NTA resin turned into something like gel structure inside the column after washing just before the elution.. Is anyone seen something like this before? What does it mean?
Thank you very much!!
Did the washing buffer contain a high concentration of a strong chaotropic agent like guanidinium thiocyanate, which may disrupt the structur of an agarose-based matrix? At any rate, I suppose that this “gellification” hampers the flow and prevents the elution with the elution buffer. You may still try to recover your protein by taking out the content of the column and performing a batch elution by resuspending It in elution buffer.
It is due to the presence of chaotropic agents .Try to check ur buffer once.
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