RP - In Solution Digest for Mass Spectrometry (MS) Analysis (»Add to Infobox)

Mann Matthias

Matthias Mann's Group

There are two major strategies for converting proteins extracted from biological material to peptides suitable for MS-based proteome analysis. The first involves solubilization of proteins with detergents, separation of proteins by SDS-PAGE (Sodium Dodecil Sulfate Polyacrylamide Gel Electrophoresis), and digestion of the gel-trapped proteins (‘in-gel’ digestion). The second is the detergent-free method comprising protein extraction with strong chaotropic reagents such as urea and thiourea, protein precipitation, and digestion of proteins under denaturing conditions (‘in-solution’ digestion). For samples containing no detergent and of low to medium complexity (<300 proteins), in-solution digestion can be used and will provide similar results to in-gel digestion. This technique can also be very useful when working with small protein amounts, since the sample recovery is better than that for in-gel digestion.

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Mann Matthias (Principal Investigator / Matthias Mann's Group)

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