Wednesday, January 18, 2017
Absorb entire cells into dried acrylamide gels for digestion!??!?
This study is paywalled...and you can't find any details until you get into the Supplemental info, but it is still neat enough that it is worth talking about!
What is it? A new and enormously efficient looking digestion method. They seriously make polyacrylamide gels dry them down completely and then add a solution containing a solution of whole cells (looks like they used non-adherent cancer cells, but I forget now).
Once the cells are inside the gel they basically treat them like a normal in-cell digestion. Compared to what they get with their normal in-gel digestion methods (and something called pro-absorb) they get massively more peptides and proteins per run on their Orbitrap Fusion.
Personally, I'd never have guessed that whole cells could be taken up like that! And, in their analysis, there were far fewer missed cleavages than the other methods they used. Now....you might argue that the total number of proteins they get out of all 3 methods seems a lot lower than what normally expect to see with more traditional methods like in-solution digestion or FASP, but this paper is clearly focused on showing the promise of this novel cell-absorption/digestion method.