Friday, January 27, 2017

Loss-less nano fractionation -- all you need is a Spider!


Sometimes you just need absolute coverage of your sample -- an MS/MS spectra (or 10) representing every theoretical peptide -- and then you find out you've only go 10 ug to work off of...which means you're pretty much stuck with long column 1-dimensional fractionation....

OR!!! You hook a SPIDER FRACTIONATOR up to an EasyNLC and get loss-less prefractionation, as described in this article in this month's MCP!


Nanoflow fraction collection?  Into 96 well plates? Virtually no sample loss? Any other cool things about it?

You don't have to buffer exchange or anything -- you can fraction collect in your first dimension with C-18 as shown above, recombine your fractions in a way that ensures a relatively equal distribution of peptides on your analytical gradient and get maximum coverage without any dumb loss steps.

Worth noting: The Spider fractionator is a beta test unit in development for commercial release by PreOmics and isn't quite available yet. You know, cause sometimes Dr. Mann gets access to stuff before the rest of us do....

How can you get away with calling this "Loss less" fractionation? Seems like an exaggeration, right? Ever fraction collected 1 (one!) ug of peptides, ran the fractions and gotten 10,000 protein groups?  I sure haven't, but...



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