Argonaute 2 (Tyr-393), Phosphospecific Antibody

Several classes of small RNAs, including short interfering RNAs (siRNAs), microRNAs (miRNAs), and Piwi-interacting RNAs (piRNAs) have been identified. MicroRNAs are about 21 nucleotides in length and have been implicated in many cellular processes such as development, differentiation, and stress response. These small RNAs function together with complexes called micro-ribonucleoproteins (miRNPs) to regulate gene expression by modulating mRNA translation or stability. Among the most important components in these complexes are argonaute proteins. There are four members in the mammalian argonaute family and only argonaute 2 (Ago2) possesses the Slicer endonuclease activity. Argonaute proteins participate in various steps of microRNA-mediated gene silencing, such as repression of translation and mRNA turnover. These activities may be regulated by cell signaling events that alter argonaute phosphorylation. EGFR phosphorylates Tyr-393 in Ago2, which reduces binding to Dicer and inhibits miRNA processing. Akt3 phosphorylates Ago2 at Ser-387 leading to reduced mRNA cleavage and enhanced translational repression.

This antibody was cross-absorbed to unphospho-Ago2 (Tyr-393), before affinity purification using phospho-Ago2 (Tyr-393) peptide. The antibody detects a 97 kDa* protein corresponding to the apparent molecular mass of phosphorylated Ago2 on SDS-PAGE immunoblots of human A431 cells that were treated with EGF.