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

Phosphorylation of Cdc25B3 Ser169 regulates 14-3-3 binding to Ser151 and Cdc25B activity.

Astuti P, Gabrielli B

Cdc25B is a major regulator of G(2) phase progression into mitosis and in recovery from a G(2) phase checkpoint arrest. The activity of Cdc25B is regulated by 14-3-3 dimer binding to a high-affinity phosphosite Ser323 and two lower-affinity phosphosites Ser151 and Ser230, which blocks substrate access to the catalytic site and, thereby, activity. Loss of 14-3-3 binding to any of ... [more]

Cell Cycle Jun. 15, 2011; 10(12);1960-7 [Pubmed: 21558810]

Throughput

  • Low Throughput

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
CDC25B CCNB1
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.

High-BioGRID
2215328

Curated By

  • BioGRID