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High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format
Authors:Cummings Edwin D  Brown Janet M  Sarva Siva T  Waldo Robert H  Hilliard George M
Affiliation:Center of Excellence in Genomics and Bioinformatics, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
Abstract:Processing multiple protein samples from polyacrylamide at significant sensitivity represents a major chokepoint for raising the success rate in high-volume protein identification projects. A multiwell filterplate method for processing proteins in polyacrylamide was optimized for sensitivity using a protein standard. The results demonstrate this process to be a reliable and reproducible method over a range of gel loadings and suitable for the identification of proteins near the threshold of silver stain. This high-throughput manual method requires a minimum of specialized equipment, and can be performed disconnected from a proteomics infrastructure for the preparation of mass spectrometry-ready samples.
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