Can I purify protein straight from the cell culture supernatant?

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Can I purify protein straight from the cell culture supernatant?

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It depends on the purification techniques. As far as we are aware, you can use techniques such as ion exchange or antibody affinity purification. However, Ni-NTA purification or other metal-affinity purification of His-tagged proteins will not work. Firstly, the cell culture medium contains other charged compounds that will compete with your protein. Secondly, the low pH (~5.5) causes the His-tag to be protonated and therefore precludes binding to the resin. You will have to change buffer to a higher pH buffer (pH 7.5+) to deprotonate the His-tag. See us on how to do this using a tangential flow filtration system and check out the protocols page.

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