Dear Dhiraj,
yes, on-column cleavage works as well quite efficiently with 3C protease.
I am performing it in a cold room (8-10C) for 1h normally reaches similar
efficiency.
Depending on your tags (if the 3C has a His-tag) you can add even more protease
1:50 then for sure 1h is more than enough.
Thanks Nikolay
I am doing on column cleavage, so I am incubating it
overnight at 4-degree C. Do you think even on column digestion is also going to
be done in one hour.
Thank you
Dhiraj
From: CCP4 bulletin board on behalf of Nikolay
Dear Dhiraj,
the 3C protease is from the Cysteine protease family.
Therefore for sure, you can add EDTA (if it is not indented for follow up IMAC
purification) and that will inhibit most of the metalloprotease. Furthermore,
you can resort to Serine type protease inhibitors like PMSF or
Hi
sorry for off topic question. does anyone know which protease inhibitors we
can include safely while cleaving with 3C protease?
Thank you
Dhiraj
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Dear Boaz
I'm replying to the bulletin, for others to also benefit from the
conversation.
I was hoping for more (positive) replies and decided I will not upgrade it
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a new SSD for a clean installation, keeping the current
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