I would like to purify a protein using the IMACT system where my protein has an intein and chitin-binding domain. I cannot use salt in my purification. The protocol says that you can use 50-1000 mM salt, but it seems like a suggestion. I was curious if anyone has had success with this purification without salt or know if I am likely to be able to get binding to the beads without salt in my buffer.
Please note that in affinity methods, salt may not only helpful for the binding process, but also to suppress non-specific binding of irrelevant proteins. Could you specify, why you cannot use salt in your purification? In most cases, it might be preferable to remove any salt after the purification, if necessary.
Thank you all for your responses! As an update, I was able to use a buffer without salt and still achieve binding to the chitin beads.
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