Catalog # AK6970

Argonaute 2 Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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.

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.

Western blot analysis of mouse recombinant Ago2 full length protein (lanes 1-4). The blot was treated with lambda phosphatase (lanes 2 & 4) then probed with rabbit polyclonal anti-Ago2 (AP5281) (lanes 1 & 2) and rabbit polyclonal anti-Ago2 (Ser-387) phospho-specific antibody (lanes 3 & 4).

Western blot analysis of human A431 cells treated with EGF (100 ng/ml for 60 min.) (lanes 1-4). The blot was treated with lambda phosphatase (lanes 2 & 4) then probed with rat monoclonal anti-Ago2 (AM5271) (lanes 1 & 2) and rabbit polyclonal anti-Ago2 (Tyr-393) phospho-specific antibody (lanes 3 & 4).



Rat monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Argonaute 2 (N-terminus) Rat mAb50 μlWB, EHu, Bv97
Argonaute 2 (a.a. 389-398) Rabbit pAb50 μlWB, EHu, Rt, Ms, Ck F97
Argonaute 2 (Ser-387), phospho-specific Rabbit pAb 50 μlWB, EHu, Rt, Ms, Ck, F97
Argonaute 2 (Tyr-393), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms, Ck, F97
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb 100 μlWB, E, ICC, IHCRb
Anti-Rat IgG Light Chain:HRP Goat pAb100 μlWB, E, ICC, IHCRt
KIT SUMMARY

The argonaute 2 (Ago2) phospho-regulation antibody sampler kit can be used to detect phosphorylation of Ser-389 and Tyr-393 relative to total Ago2 expression levels. The kit includes rabbit polyclonal phospho-specific antibodies to Ser-389 and Tyr-393, as well as rat monoclonal and rabbit polyclonal antibodies for detection of total Ago2. The kit also includes anti-Rabbit Light Chain specific:HRP and anti-Rat Ig:HRP secondary reagents for detection of antibodies in western blot, ELISA, or immunocytochemistry.