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imidazole in protein purification - (Nov/11/2005 )

why does the 1st buffer in his-tagged protein purification contain a little bit of imidazole? please help me !!

-biobuddy-

To wash off contaminants which may bind non-specifically to the column. Ideally your his tagged protein will have a much higher affinity to the resin then non his tagged proteins, so it will remain bound in the presense of low imidazole while others get washed off.



QUOTE (biobuddy @ Nov 11 2005, 09:22 AM)
why does the 1st buffer in his-tagged protein purification contain a little bit of imidazole? please help me !!

-Cole-

according to Qiagen, imidizole also increases the solubility of your rProtein, but I'm not sure why that is the case...if I remember correctly, imidizole reduces proteins; perhaps this helps to break up aggregates, which will often form when you've got tons of rProtein hanging around in the cell...is this right? anyone? anyone?

-aimikins-