Can 1% tritonx-100 remain the integrity of the complex - (Jan/12/2007 )
Use 1% triton X-100 th lyse the cell, want to recover a tagged membrane protein and it's binding parters. Will the protein complex remain together in the presence of 1% triton?
I would say the binding partners would most likely be dissociated. Try lower % of triton. We use 0.1% triton in the extraction buffer.
If u r lucky, may b u could still detect the binding partners.
Some maybe useless experience...
I once detected intertraction of a binding partner of a TM protein with both 2% NP40 and 0.5% NP40..... HOWEVER, when I repeated the experiment, even 0.5% NP40 didn't work.... I have been still struggling with lysis buffer for months.... (I haven't try 0.1% yet...)
i use to do my coIP in 0.05 to 0.1% NP40
1% seems quite high to me to maintain complex association.
I think it may be ok for polyproteins (like LDH for ex) but protein partners are quite dissociated.
What is the salt concentration used?
I do 0.05% NP40 and 300mM KCl.
1% triton x-100 will lyse the cell but membrane protein complexes may be disintegrated; there arises another problem in Co-IP if conc of triton is above CMC so that protein complexes are trapped in micells