So class, I have a quick question about Co-IP detection. In the case of MULE, my antibody would bind EGFR (bound to MULE), then wash out unbound crap to have Ab-EGFR-MULE. Under normal circumstances, I would just run this on a gel and do a western blot against MULE to confirm binding, but without the MULE Ab this doesn't work. Is there another way of detecting secondary targets in Co-IP? otherwise I'm just thinking of doing a different binding assay, but I'm curious about how the MULE discoverers did it.
Also, someone talked about using RT-PCR from MULE RNA to isolate the gene. But given the similarities with RAS, wouldn't we end up with a mixture of the two? I'm thinking in terms of primers, because the only differences are in UTRs, which you won't have in the cDNA... so how can the primers be specific for just MULE?