Affinity Capture-Western

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner identified by Western blot with a specific polyclonal antibody or second epitope tag. This category is also used if an interacting protein is visualized directly by dye stain or radioactivity. Note that this differs from any co-purification experiment involving affinity capture in that the co-purification experiment involves at least one extra purification step to get rid of potential contaminating proteins.

Publication

Composition and Regulation of the Cellular Repertoire of SCF Ubiquitin Ligases.

Reitsma JM, Liu X, Reichermeier KM, Moradian A, Sweredoski MJ, Hess S, Deshaies RJ

SCF (Skp1-Cullin-F-box) ubiquitin ligases comprise several dozen modular enzymes that have diverse roles in biological regulation. SCF enzymes share a common catalytic core containing Cul1â‹…Rbx1, which is directed toward different substrates by a variable substrate receptor (SR) module comprising 1 of 69 F-box proteins bound to Skp1. Despite the broad cellular impact of SCF enzymes, important questions remain about the ... [more]

Cell Nov. 30, 2017; 171(6);1326-1339.e14 [Pubmed: 29103612]

Throughput

  • Low Throughput

Additional Notes

  • An interaction with Cul1 was detected using an anti-phospho-Akt antibody. It is not clear which Akt isoform (Akt1, Akt2, or Akt3) this detects. Therefore all 3 have been entered.

Curated By

  • BioGRID