High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format.
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.
Cummings
Edwin D
ED
Center of Excellence in Genomics and Bioinformatics, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
Brown
Janet M
JM
Sarva
Siva T
ST
Waldo
Robert H
RH
Hilliard
George M
GM
D000180:Acrylic Resins / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods* / Q000592:standards; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods* / Q000592:standards; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D014357:Trypsin / Q000737:chemistry; D000099166:Polyacrylamides
D000180:Acrylic Resins; D011506:Proteins; D012710:Serum Albumin, Bovine; D014357:Trypsin; D000099166:Polyacrylamides