Optimized Affinity Capture of Yeast Protein Complexes

Cold Spring Harb Protoc. 2016 Jul 1;2016(7). doi: 10.1101/pdb.prot087932.


Here, we describe an affinity isolation protocol. It uses cryomilled yeast cell powder for producing cell extracts and antibody-conjugated paramagnetic beads for affinity capture. Guidelines for determining the optimal extraction solvent composition are provided. Captured proteins are eluted in a denaturing solvent (sodium dodecyl sulfate polyacrylamide gel electrophoresis sample buffer) for gel-based proteomic analyses. Although the procedures can be modified to use other sources of cell extract and other forms of affinity media, to date we have consistently obtained the best results with the method presented.

MeSH terms

  • Affinity Labels / metabolism
  • Chromatography, Affinity / methods*
  • Electrophoresis
  • Macromolecular Substances / isolation & purification*
  • Multienzyme Complexes / isolation & purification*
  • Proteome / analysis
  • Saccharomyces cerevisiae / chemistry*
  • Saccharomyces cerevisiae Proteins / isolation & purification*


  • Affinity Labels
  • Macromolecular Substances
  • Multienzyme Complexes
  • Proteome
  • Saccharomyces cerevisiae Proteins