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

Immunoprecipitation techniques for the analysis of transcription factor complexes.

Klenova E, Chernukhin I, Inoue T, Shamsuddin S, Norton J

Interactions among transcription factors can be detected and analyzed by a variety of in vitro and in vivo approaches. In many studies, the existence of putative interactions among transcription factor partners is initially established from yeast two-hybrid screening and in vitro protein association analysis. The ability to detect candidate interacting proteins in coimmunoprecipitates from cell lysates provides an important criterion ... [more]

Methods Mar. 01, 2002; 26(3);254-9 [Pubmed: 12054881]

Throughput

  • Low Throughput

Curated By

  • BioGRID