Exploring human plasma proteome strategies: high efficiency in-solution digestion protocol for multi-dimensional protein identification technology |
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Authors: | Ru Qinhua Cindy Zhu Luwang Andy Katenhusen Richard A Silberman Jordan Brzeski Henry Liebman Michael Shriver Craig D |
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Affiliation: | Windber Research Institute, PA 15963, USA. c.ru@wriwindber.org |
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Abstract: | 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. |
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