Important Notice: Our web hosting provider recently started charging us for additional visits, which was unexpected. In response, we're seeking donations. Depending on the situation, we may explore different monetization options for our Community and Expert Contributors. It's crucial to provide more returns for their expertise and offer more Expert Validated Answers or AI Validated Answers. Learn more about our hosting issue here.

Can I purify protein straight from the cell culture supernatant?

0
Posted

Can I purify protein straight from the cell culture supernatant?

0

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.

Related Questions

Thanksgiving questions

*Sadly, we had to bring back ads too. Hopefully more targeted.