High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format

J Proteome Res. 2007 Apr;6(4):1603-8. doi: 10.1021/pr060472y. Epub 2007 Mar 17.

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.

MeSH terms

  • Acrylic Resins / chemistry
  • Amino Acid Sequence
  • Animals
  • Cattle
  • Molecular Sequence Data
  • Peptide Mapping / methods*
  • Peptide Mapping / standards
  • Proteins / chemistry
  • Proteins / isolation & purification*
  • Proteomics / methods*
  • Proteomics / standards
  • Serum Albumin, Bovine / chemistry
  • Serum Albumin, Bovine / isolation & purification
  • Trypsin / chemistry

Substances

  • Acrylic Resins
  • Proteins
  • Serum Albumin, Bovine
  • polyacrylamide
  • Trypsin