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Protein interaction with His-tagged protein


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#1 Ecolina20

Ecolina20

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Posted 29 June 2011 - 11:14 PM

Hi,

I want to find interaction partners of my His-tagged protein in the mammalian cell line that I am expressing the protein in (and differences with and without stimulation). Knowing that in during purification of His-tagged proteins other undesired but probably interacting proteins might co-elute, I was hoping to use that phenomenon to find potential interaction partners. Normally manufacturer`s protocols will describe His-tag pull down assays using lysates of different cells expressing the different proteins, with wash steps in between. Since both prey and bait are in the same cell in my case I thought I could probably modify the protocol (e.g. washing) accordingly.

Does anybody have experience with that or ideas what to try?

Thanks!




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