Dear all,
I agree with all said so far:
Here are some points from my experience and discussion on this topic.
Most of the lab's due use TEV or 3C, depending on the cleavage site in their
constructs and make their purification strategy work with what they have.
Here are some points from my side:
In my experience, 3C can partially cut TEV sites as well. If using
sequentially, it is best to plan to use the TEV first.
> On Dec 7, 2022, at 2:42 PM, Lau Kelvin
> <5aaf8435dbef-dmarc-requ...@jiscmail.ac.uk> wrote:
>
> I agree. I think it depends on the lab and vectors and personal
I agree. I think it depends on the lab and vectors and personal preference.
But David, have you used them sequentially? I have once tried to make a
His-GST-ENLYFQ-3C construct, and I found that it was self cleaving during
expression. However I have never tried to replicate the results in vitro.
HRV3C cut of an N-terminal tag usually leaves GP at the N-terminus. TEV usually
leaves G/S. Structural biologists may prefer TEV. However, HRV3C has more
robust activity than TEV. HRV3C is a better choice for removal of C-terminal
tags.
To save precious time, Accleagen has Turbo3C
I believe HRV3C might be more efficient if you need to cleave in the absence of
reducing agents (e.g. extracellular proteins).
Best regards,
Cyprian
Cyprian Cukier, PhD
Selvita S.A.
From: CCP4 bulletin board On Behalf Of Crissy L Tarver
Sent: Wednesday, December 7, 2022 9:38 PM
To:
We use the HRV3C protease site frequently in our lab. It provides better
on-bead/column cleavage than TEV. We have a plasmid as we make 3C protease in
our lab and keep stocks in our -80C. I can send you our protocol if you wish.
Crissy
Crissy L Tarver, PhD
Department of Structural Biology
Hi Gloria,
Both can be made very easily in E.coli.
Both are active at 4°C, but especially 3C, I think.
I have plasmids for both somewhere in the freezer (you might find someone
closer to you who can send HRV3C, but if you cannot, let me know off list).
I don't see any particular benefit of one
Hello my fellow structural biologists, I am contemplating why some choose
the HRV3C protease site over TEV for their fusion proteins. Does anyone
know? Can HRV3C be made easily in homelab? Does anyone have a plasmid?
Thank you, G
The Hauptman-Woodward Medical Research Institute (HWI)
(https://hwi.buffalo.edu) seeks an established investigator to join the
scientific efforts at the Institute. Applications in the broad areas of
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seeks exceptional
Hi Yufeng,
You can use NVH Oil from jenabiosciences.com as alternative, it is a
high viscosity oil.
You can also use STP oil tratement. It has been suggested by Vic Young
on the Bruker mailing list and approved by several users there. Joe
Reibenspiess put it on his web site for utilities
Hi there,
I just noticed that Hampton Research discontinued Paratone-N and now recommends
Santovac cryo oil. Has anyone tried Santovac and can comment on how it compares
to Paratone? We had some previous success of using paratone for crystal
annealing and felt it was a good cryoprotectant
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