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Updated: Apr 4, 2026

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans
Published on: May 23, 2020
Biochemical isolation of Argonaute protein complexes by Ago-APP
Judith Hauptmann1, Daniel Schraivogel1, Astrid Bruckmann1
1Biochemistry Center Regensburg, Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany;
Abstract:
During microRNA (miRNA)-guided gene silencing, Argonaute (Ago) proteins interact with a member of the TNRC6/GW protein family. Here we used a short GW protein-derived peptide fused to GST and demonstrate that it binds to Ago proteins with high affinity. This allows for the simultaneous isolation of all Ago protein complexes expressed in diverse species to identify associated proteins, small RNAs, or target mRNAs. We refer to our method as "Ago protein Affinity Purification by Peptides" (Ago-APP). Furthermore, expression of this peptide competes for endogenous TNRC6 proteins, leading to global inhibition of miRNA function in mammalian cells.

