Need to teach a class about report ion quantification? Holy smokes, Yavin Raunivar and John Yates just put together your lesson plans in this new review in JPR.
Its thorough, up-to-date, and shockingly concise considering the history, reagents and methodologies described. Even if you've done these experiments for years and with different instruments there are still some great insights here!
There is a great section on doing PTM quantification with reporter ions (very phospho-centric) that brings up a really interesting methodology (reversed ammonia gas spray across the front of the instrument (?what?!? I gotta read that) that boosts TMT-phospho IDs (??again, no idea!!).
The highlight in this, for me, is a concept I've never even considered and I feel really dumb for not having come up with myself. TARGETED ANALYSIS with reporter ions. We're getting more targeted all the time, especially with new high-certainty LC-MS methods like PRM (parallel reaction monitoring). These let us look at hundreds of peptides in a pathway and each MS/MS scan gives us a ton of confirmational data that we're looking at the right target. But what if we multiplexed it with TMT-10? Then you get the sensitivity of the targeted approach and the certainty from the PRM and you can also get relative quan from up to 10 patients at once!!!
Sorry, my mind is kind of blown. I'd better finish this coffee and get to work....
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