Dear Chen,
The two proteins try to be crystallized, are exactly the same construct with the two proteins have been tested in co-IP?
A his tag may affects binding.
The two proteins have been tested in co-IP were expressed in E.coli?
Post translation modification for sure effect function and thus binding
The PUll-down assay or gel fitration was taken place at the same or similar pH and salt concentrations with those of co-IP ?
Usually in gel filtration medium or relative high salt concentration is used
George
-----Original Message-----
From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of <Xia Chen>
Sent: Thursday, January 14, 2016 1:34 PM
To: [log in to unmask]
Subject: [ccp4bb] how to enhance the complex interaction(weak)
Dear every one:
nearly, I have try to crystalize two protein complex. Although we get know from the cooperator that two protein have interaction just for Co-immunoprecipitation (IP). when I express them in E.coli. regardless of PUll-down assay or gel fitration, we can't dectect the interaction. I want to find some ways to enhance their interaction so that I can crystalize the complex.
Chen Xia, Ph.D
School of Life Sciences
Tianjin University
92 Weijin Road, Nankai District, Tianjin 300072, P.R.China
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