Publication

Affinity proteomics to study endogenous protein complexes: Pointers, pitfalls, preferences and perspectives

LaCava, J., Molloy, K. R., Taylor, M. S., Domanski, M., Chait, B. T. & Rout, M. P., Mar-2015, In : Biotechniques. 58, 3, p. 103-119 14 p.

Research output: Contribution to journalReview articleAcademicpeer-review

  • John LaCava
  • Kelly R. Molloy
  • Martin S. Taylor
  • Michal Domanski
  • Brian T. Chait
  • Michael P. Rout

Dissecting and studying cellular systems requires the ability to specifically isolate distinct proteins along with the co-assembled constituents of their associated complexes. Affinity capture techniques leverage high affinity, high specificity reagents to target and capture proteins of interest along with specifically associated proteins from cell extracts. Affinity capture coupled to mass spectrometry (MS)-based proteomic analyses has enabled the isolation and characterization of a wide range of endogenous protein complexes. Here, we outline effective procedures for the affinity capture of protein complexes, highlighting best practices and common pitfalls.

Original languageEnglish
Pages (from-to)103-119
Number of pages14
JournalBiotechniques
Volume58
Issue number3
Publication statusPublished - Mar-2015
Externally publishedYes

    Keywords

  • protein complex, protein purification, affinity, proteomics, interactomics, NUCLEAR-PORE COMPLEX, PURIFICATION-MASS-SPECTROMETRY, INDUCIBLE GENE-EXPRESSION, EPSTEIN-BARR-VIRUS, MAMMALIAN-CELLS, SACCHAROMYCES-CEREVISIAE, MOLECULAR ARCHITECTURE, BAC TRANSGENEOMICS, ENHANCED DETECTION, RAPID PRODUCTION

ID: 117137265