Enzyme Purification Questions

Hi there,
I have a feeling that many more people have experience with protein expression (even GFP) than protein purification, could people maybe post resources/guides that explain the process more thoroughly? Something like the review on e.coli expression from the open bio-economy ENZYME EXPRESSION GUIDE? Or any other guide? Something wide, and maybe also something more specific to His tagged proteins?

There has also been a lot of discussion about the nickel columns and getting them, could somebody advise which ones are the best to buy?

Thanks a lot

Hey Guy!
Nice to meet you here :slight_smile: I found the GE Healthcare guides most helpful: https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/General_Information/1/ge-recombinant-protein-purification.pdf
I also use the Chelating Sepharose Fast Flow: https://static.fishersci.eu/content/dam/fishersci/en_EU/lifescience/doctechnical/proteomics/GE_Data%20file_Chelating%20Sepharose%20Fast%20Flow.pdf, because I got some leftovers of a working group I know. It’s pretty expensive, though.
But you can load other Kations than Nickel and clean/strip & reuse it.

Hey you all,
I did a little Enzyme Purification Playlist on Youtube with some resources, I consider helpful to get started. Here you go: https://www.youtube.com/watch?v=nY8jW9XbYSY&list=PLNFCojEhItgB94IrGpXeSAZEDpAqvzbHy