Saturday, September 3, 2016
High pH reverse phase StageTip FTW!
Need a quick method to boost your membrane proteomics coverage?
At first glance this paper might seem a little boring or obvious, but I only know one group that uses techniques like this and they've never published it and I'd never guessed it would be as powerful as this new paper suggests.
The idea is that they solubilize with SDS-PAGE buffers (which....despite tons of work on other techniques...is still probably the best technique for high membrane proteome coverage), run a gel and gel extract. I've seen some groups run for only a very short time and then only cut out the very very top piece of the gel (right below the stacking) and digest it out. There is still a TON of stuff in there.
This group (sorry, first author, your name is way too long for me to write it out here et al.,) takes it another step forward, taking the gel slice StageTip high PH reverse phase fractionating it and walk away with a huge number of membrane proteins and peptides with typical membrane spanning domains from a tiny amount of membrane starting material.
Simple and cheap boost to membrane proteomics coverage? Sign me up!