Wednesday, August 5, 2015
Massively upgrade your FASP with MStern blotting!
I admit it. I HATE western blots. There's nothing like coming in with data from one of the most sensitive/accurate analytical devices every conjured up by physics and being told to "validate" these observations with funny colored rabbit blood.
Fortunately for us, MStern blotting has nothing at all to do with Western blots at all, except that it makes use of PVDF membranes to massively speed up our ability to do FASP-style digestions.
I stole the figure above from this paper by Sebastien Berger et. al., that is currently open access at MCP here. (oh, some Steens were involved here as well!)
The idea is this: you can do FASP in 96 well plates (a very nice JCVI paper can be found on this blog). The problem is that it takes a long time with most filters. By substituting PVDF membranes for MW cutoff filters they were able to get great digestions an cleanups in much faster times using speeds that are compatible with 96-well plates.
Awesome? Absolutely. And not just because I don't have to do a western blot! Definitely check this one out!