Hi everybody!
Im just started in coimmunoprecipitation world.
Apparently it works fine because I can detect the protein A and the protein B separate, and the negative controls are ok but with some background.
My problem is when I have A and B proteins together, I detect A and B when I coimmunoprecipitate with B, but when I coimmunoprecipitate with A I only detect A and not B (big and glicosilated protein).
I think they are interacting but I dont know why I can see it only in one way.
Ive tried to put more lysate when I immunoprecipitate with B, but I hadnt any improvement.
I dont know why it occurs, please if somebody can help me Ill be very grateful.
Thanks
Esther

Hi,
Congratulation and welcome to the Co-IP world.
One suggestion is to try to IP protein A with another antibody. Or try to detect Bwith a different Ab.
Sometimes the epitope hides.
Try another lysis buffer.
Guy