This topic contains 2 replies, has 2 voices, and was last updated by Eyad Fansa 7 months, 2 weeks ago.
December 7, 2018 at 13:31 #1997
this is Eyad from the DPF facility in MPI-Dortmund. I was wondering whether someone have a good experience with the purification of human Caspase-1 from e.coli.
I have the following three ideas:
1- Separate expression of the p20 and p10 subunits followed by refolding from enclusion bodies.
2- Repalcement of the autoproteolysis sites D119 and D279 by thrombine cleavage sites.
3- Purify the full lenght protein in the prescence of Caspase inhibitor followed by inhibitor removal and autoactivation of the protein.
I appreciate your comments or suggestions.
December 7, 2018 at 13:52 #1998
did it before. Express full-length, refold and concentrate really, really high, this force autoactivation. After dilution you can polish with a final purification step.
December 10, 2018 at 14:24 #1999
Many Thanks, Eyad
You must be logged in to reply to this topic.