Tags for protein purification
PURUSHARTH RAJYAGURU
Monday, 20 August 2007 20:00 UTC
Hello everybody,
I have a bunch of proteins with different tags as His, GST, FLAG . All these proteins are a part of a complex. I am now looking for a new tag (besides the above ones) to put on the protein and this one will be used to pull down the entire complex? I need to use bacteiral expression vectors.
Can anybody suggest me on possible good alternative tags?
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Replies
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Hi,
I recommend c-myc tag or even better tandem affinity purification tag. Unfortunately, I can’t provide you with any plasmids since it is a long time now I left the field.
Good luck!!
Marek -
also GFP tag exists in mammalian cells.
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sorry i didn’t find how to edit my previous post.
Well HA tag exist also. The efficiency of recovery is ok, but it’s less specific. Agarose coupled anti HA is available from sigma and other brands -
Thanks for the responses but as I have mentioned I need to use E. coli expression system and I am not sure if c-myc, HA etc would work in bacteria.
Any comments? -
Hi,
c-myc, GFP, HA will work in E.coli as well. HA tag usually gives higher background after purification.
Best
Marek -
I found this website while I was looking for protein purification protocols to commission:
http://wolfson.huji.ac.il/purification/Expression_Systems.html.Maybe it will be useful to you as well?
Best of luck!
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You can use S-protein tag. pET30b vector from Novagen has this tag. As far as I know, S tag and His tag are in tandem in this vector, though. But this vector has such features that allow you to rid of His or His/S tags from the protein after purification. Novagen sells S tag purification kits.
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Hi purusharth, this is rahul here…I hope u remember, well u can go to a site of Arabidopsis Biological Resource Center..They have quite a few vectors having differential tags such as MYC, HA for the bacterial expression system and that too for just 4 USD each vector..Order from their…U will definately find some of ur interest. Tk care
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