RP - In Gel Digestion 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). The one dimensional SDS-PAGE can be used not only to reduce sample complexity, but also to remove sample contaminants as detergents, salts, DNA and other non-protein compounds. However, the sample recovery in in-gel digestions is likely much less than in in-solution digestions, so this technique is more indicated for very complex samples (>300 proteins) where sample amount is not limited.

MEMBERS (1/1)

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

DOWNLOADS (1/1)