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

Targeted proteomic analysis of 14-3-3 sigma, a p53 effector commonly silenced in cancer.

Benzinger A, Muster N, Koch HB, Yates JR, Hermeking H

To comprehensively identify proteins interacting with 14-3-3 sigma in vivo, tandem affinity purification and the multidimensional protein identification technology were combined to characterize 117 proteins associated with 14-3-3 sigma in human cells. The majority of identified proteins contained one or several phosphorylatable 14-3-3-binding sites indicating a potential direct interaction with 14-3-3 sigma. 25 proteins were not previously assigned to any ... [more]

Mol. Cell Proteomics Jun. 01, 2005; 4(6);785-95 [Pubmed: 15778465]

Throughput

  • Low Throughput

Additional Notes

  • Tandem affinity purification (TAP) and the multidimensional protein identification technology (MudPIT)

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
SFN AJUBA
Affinity Capture-MS
Affinity Capture-MS

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 is identified by mass spectrometric methods.

Low-BioGRID
244921

Curated By

  • BioGRID