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

A Distinct Role of Riplet-Mediated K63-Linked Polyubiquitination of the RIG-I Repressor Domain in Human Antiviral Innate Immune Responses.

Oshiumi H, Miyashita M, Matsumoto M, Seya T

The innate immune system is essential for controlling viral infections, but several viruses have evolved strategies to escape innate immunity. RIG-I is a cytoplasmic viral RNA sensor that triggers the signal to induce type I interferon production in response to viral infection. RIG-I activation is regulated by the K63-linked polyubiquitin chain mediated by Riplet and TRIM25 ubiquitin ligases. TRIM25 is ... [more]

PLoS Pathog. Aug. 01, 2013; 9(8);e1003533 [Pubmed: 23950712]

Throughput

  • Low Throughput

Ontology Terms

  • cell type: hek-293ft cell (BTO:0005029)
  • cell type: huh-7 cell (BTO:0001950)

Additional Notes

  • HEK293FT cells were stimulated with 0.8 ug of HCV double-stranded RNA (HCV RNA)followed by immunoprecipitation with an anti-RIG-I mAb (Alme-1).
  • SeV infection induced the association of endogenous RIG-I with TRIM25 and TBK1, whereas HCV replicons failed to induce the association

Curated By

  • BioGRID