Replies to post #160719 on Biotech Values

[poorgradstudent]

ARQL:

But my question is whether looking for the c-Met unmutated mRNA will find heavily mutated c-Met (as many consitutively active c-Mets are). I.e. I believe that the two assays they did would miss a high fraction of constitutively actived c-Met. But that said, the reason I asked for vin (or you) to look is that I don't know the precise limits of the two assays they used - so I was looking for someone to comment on that.

Right, and the antibodies they used for the immunoprecipitations were polyclonal, so they would not be meaningfully effected by point mutations in the protein. So I think it is not necessary to worry about their assay not capturing c-Met mutants.

[vinmantoo]

iwfal,

Good eye, and I agree with your assessment. Sorry poorgradstudent but I disagree with you. In the paper Iwfal cited, they don't even tell us which primers they used for detecting c-MET RNA. The 2nd paper, the one poorgradstudent cited, did list the primers for assessing c-MET RNA but they only used a single primer pair, which is very sloppy. If one of those regions was deleted in the genome, or mis-spliced so absent, so they could have easily missed identifying the mRNA of a truncated gene. The paper iwfal sited is vague about the anti-MET antibodies, but figure 1A shows it as CT which I assume is the the C-terminal domain of c-MET and the blots in figs 1E & 6A indicate this as well. If that C-terminal domain was missing in TOV cells, and one of the primers was from that region, you could miss both the RNA and protein.