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Technical Data Sheet

AX5295

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phospho-Argonaute 2 (Ser-387)
Blocking Peptide
Price
Size

$55
50 μg

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Application
Dilution


Blocking
1:1000


ELISA
50 ng/well



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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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.


Background References
Peters, L. & Meister, G. (2007) Mol Cell 26:611.
Filipowicz, W. et al. (2008) Nat Rev Genet. 9:102.
Horman, S.R. et al. (2013) Mol Cell 50:356.
Shen, J. et al. (2013) Nature 497:383.
Sequence
Phospho-Ago2 (Ser-387) synthetic peptide corresponding to amino acid residues surrounding serine 387 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 the homology surrounding the site is low between Ago2 and these two family members.
Buffer and Storage
Blocking Peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide.
Store at –20°C. Stable for 1 year.
Specificity
The peptide is specifically recognized by anti-Argonaute 2 (Ser-387) phospho-specific antibody (AP5291) in ELISA, and has been shown to block the reactivity of AP5291 in Western blot. In addition, the peptide is recommended for use in blocking AP5291 reactivity in immunocytochemistry.
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