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

Binding of alphaII spectrin to 14-3-3beta is involved in NCAM-dependent neurite outgrowth.

Ramser EM, Buck F, Schachner M, Tilling T

Members of the 14-3-3 protein family have been implicated in neuronal migration, synaptic plasticity and learning. Using affinity chromatography followed by mass spectrometry analysis, we show here that the cytoskeletal protein alphaII spectrin is a novel ligand of 14-3-3beta. We found that 14-3-3beta interacts with alphaII spectrin via the mode 2 14-3-3 binding motif RLIQS(1302)HP. Binding required phosphorylation of Ser(1302) ... [more]

Mol. Cell. Neurosci. Sep. 01, 2010; 45(1);66-74 [Pubmed: 20598904]

Throughput

  • Low Throughput

Additional Notes

  • figure 6.

Curated By

  • BioGRID