I'm trying to do a co-IP looking at a GPI-anchored protein and a receptor. I want to be careful while selecting a lysis buffer, to ensure that I don't disrupt the interaction. Can anyone recommend a protocol for membrane protein interactions and the best lysis buffers? Thanks!
CHAPS, CHAPSO and digitonin should work well for this purpose. I remember there were some very successful results Preseniln-1 using these reagents. Good guide on selection of detergents can be found here: http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/2/biofiles_issue3_3.pdf
we are using TnT reticulocyte lysate system when we just want to know if there is a Interaction or not, thus we couple magnetic beads with suitable ligands and then incubate directly with expressed receptor protein in TnT, afterwards with the second protein in TnT . It works very well
I copy answer of Atur Udgata to similar question :
"If you are looking for co-IP of membrane proteins try using NP-40 as a detergent rather than triton-x100. Works better in my experience. Resuspend the cells in lysis buffer containing np-40 and keep them on a rotator at 4 degrees for 30 minutes before pelleting them down. Good luck !"
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