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

Interaction of synaptophysin with the AP-1 adaptor protein gamma-adaptin.

Horikawa HP, Kneussel M, El Far O, Betz H

Synaptophysin is one of the most abundant proteins of the synaptic vesicle membrane. Here, we selected the cytoplasmic carboxyterminal region of synaptophysin to search for interacting proteins by using the yeast two-hybrid system. This identified gamma-adaptin, a component of the AP-1 adaptor complex, as a synaptophysin binding protein. An anti-synaptophysin antibody coimmunoprecipitated gamma-adaptin from brain extracts, and immunocytochemistry disclosed a ... [more]

Mol. Cell. Neurosci. Nov. 01, 2002; 21(3);454-62 [Pubmed: 12498786]

Throughput

  • Low Throughput

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
AP1G1 SYP
Two-hybrid
Two-hybrid

Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.

Low-BioGRID
-
SYP AP1G1
Two-hybrid
Two-hybrid

Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.

Low-BioGRID
-

Curated By

  • BioGRID