Monday, February 27, 2017
A better phosphotyrosine enrichment protocol?
I saw a killer talk last week, centered on the ongoing work detailed in part in this recent paper. The talk was an overview of the years-long work trying to understand phosphotyrosine signaling in the most nefarious forms of breast cancer. I took a lot of notes -- but at the top of my page where I scrawl all my questions (and often draw skateboarding pugs) -- I ended up circling and asking something like:
"Are the antiphosphotyrosine antibodies any better than they were?" Cause...I very energetically followed the original FACE protocol within weeks of this paper coming out....and, don't get me wrong, it worked -- but that antibody pulls down a lot of stuff that ain't got a phosphotyrosine on it! Which is...frustrating...because everyone wants a bunch of phosphorylated tyrosines from you and not excuses.
Dr. Wu said it's still the case -- you pull down a lot of other things. Bummer.
And this is why I'm excited about this paper!
Here they don't try to improve the antibodies. If it was easy, we know somebody would have already done it, right? They mix up the protocol -- adding in steps that negate weak binders and they demonstrate markedly better enrichments. Hey -- whatever it takes -- cause the pY seem to be the most interesting ones!