Exploring human plasma proteome strategies: high efficiency in-solution digestion protocol for multi-dimensional protein identification technology.
Multi-dimensional protein identification technology (MudPIT) is becoming a prevalent proteomic approach due to its high-throughput separations and accurate mass detection. Prior to MudPIT analysis, complicated samples required in-solution digestion. Unlike in-gel digestion, in which enzymes work on just a few proteins, in-solution digestion involves simultaneous digestion of hundreds or thousands of proteins. In-solution digestion protocols must therefore be very efficient. Few investigations have evaluated the efficiency of in-solution digestion protocols. The present research compared three such protocols. Results suggest that a protocol utilizing trifluoroethanol (TFE) as denaturant is most efficient.
Ru
Qinhua Cindy
QC
Windber Research Institute, PA 15963, USA. c.ru@wriwindber.org
Zhu
Luwang Andy
LA
Katenhusen
Richard A
RA
Silberman
Jordan
J
Brzeski
Henry
H
Liebman
Michael
M
Shriver
Craig D
CD
D001798:Blood Proteins / Q000235:genetics / Q000302:isolation & purification; D020869:Gene Expression Profiling; D006801:Humans; D020543:Proteome; D012996:Solutions
D001798:Blood Proteins; D020543:Proteome; D012996:Solutions