I am looking to perform a co-IP; however, I am using a very small amount of tissue to begin with from region specific microdissection in mice (2-3 300um sections per animal). I am concerned that there will not be a sufficient amount of protein and/or the sample will be dilute. Are there any best practices for performing these types of experiments when the yield is low? I know one way would be to collate samples across animals, but this is something I am trying to avoid.
Thanks in advance!
no tissue co-IP experience.
however, if the antigen can be directly detected by western blot,
the co-IP should works.
It depends on whether you are using a cross-linking procedure and pulling it down or using target pull down. I would first test a control tissue of the same size on a Western to get a feel for abundance of my target protein. If there is considerable staining in WB then I would expect less intensity from co-ip and perhaps lesser for partners if it is a multi-partner protein so ratio of abundance will rely on % bound. For retrieval of protein partners I find cross-linking IP (CLIP) better.
I agree with Rabeah, have a quick look at your blot from control tissue, at least can give your general impression of your chance of pull down the target and immunoblot them.
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