FOR immunoprecipitation in plant , flag tag or His tag (View forum version)



ningjing

Posted 23 January 2011 - 12:41 PM

I want to immunoprecipitation my protein from transgenic plant. At begining, I prepare to fuse my protein with 3xFLAG tag, then use A2220 (ANTI-FLAG M2 Affinity Gel) to do the immunoprecipitate (there are a lot of paper do immunoprecipitation as I described ). But several day ago, someone suggest me to fuse my protein with 6XHis tag,than use Ni-NTA for the immunoprecipitation. (I can't find paper do like this)
My question is which one is better and why?

ningjing

Posted 26 January 2011 - 07:30 AM

up

antibodymania

Posted 26 January 2011 - 07:52 AM

Apparently His-tags are suited for affinity techniques versus FLAG-tags are better for chromatography [http://en.wikipedia....ki/Protein_tag]

I've been told the same thing, to use Ni-NTA resins because of its higher binding capacity.