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Hi,
I'm new to the area of IP and western blotting. I'm trying to look at the interaction between two proteins and after many attempts have been able to get the conditions to work for the IP/co-IP. I have noticed that once I run my IP samples and freeze them at -80C or -20C and when I try to reuse them to run the blots ( after boiling them again @ 100C for ~ 5min) that my signal has been going away. Both in the IP samples and the control in both IP and Co-IP situations. .
is it that my protein is getting degraded? I was wondering if there is any advice how to get around this issue.
Since I need to blot for several other proteins I can't run all gels at the same time.
