Argonaute 2 (Tyr-393), phospho-specific polyclonal, anti-human, mouse, rat, chicken, fish

Background:
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.

Immunogen: Phospho-Ago2 (Tyr-393) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding tyrosine 393 in human Ago2. This peptide sequence is highly conserved in mouse, rat, chick, and fish Ago2. The site is also conserved in Ago1 and Ago4, but not in Ago3.

Specificity: This antibody was cross-absorbed to unphospho-Ago2 (Tyr-393), before affinity purification using phospho-Ago2 (Tyr-393) peptide (without carrier). 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. This reactivity is not observed after lambda phosphatase dephosphorylation.

Buffer/Storage:
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.