I have isolated proteome of a bacteria and I want to isolate and purify particular proteins from them. I am not able to understand how to design a protocol for isolation and purification of them. I want to know on what basis I should isolate what kind of protein. I have already done SDS PAGE, but was unable to design a protocol for the work ahead.
You'll probably need to do some column chromatography or HPLC.
If you know the protein sequences that you want to purify, you can determine pI values for the proteins and separate by ion-exchange chromatography and then separate by size exclusion. That should enrich for your proteins. Check the fractions by western blot, then scale up to purify larger amounts. If there are good antibodies against these proteins, you can use affinity chromatography to enrich.
Good luck!
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