Cell Signaling Technology

Product Pathways - Translational Control

SignalSilence® DDX5 siRNA II #8627

Applications Reactivity
Transfection H (M)

Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® DDX5 siRNA I #8626 (+) or SignalSilence® DDX5 siRNA II (+), using DDX5 (D15E10) XP® Rabbit mAb #9877 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The DDX5 (D15E10) XP® Rabbit mAb confirms silencing of DDX5 expression, while the β-Actin (13E5) Rabbit mAb is used as a loading control.

Description

SignalSilence® DDX5 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit DDX5 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM DDX5 siRNA II 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

DDX5 (DEAD box polypeptide 5), also known as p68, was first identified as a 68 kDa nuclear protein with similarity to translation initiation factor eIF-4A (1). DDX5 is a member of the DEAD box family of putative RNA helicases, defined by the presence of a conserved DEAD (Asp-Glu-Ala-Asp) motif that appears to function primarily in the regulation of RNA secondary structure. DDX5 exhibits ATP-dependent RNA helicase activity (2) and has been identified as a critical subunit of the DROSHA complex that regulates miRNA and rRNA processing (3,4). DDX may also regulate mRNA splicing (5) and has been shown to interact with HDAC1, where it can regulate promoter-specific transcription (6). DDX5 interacts with a diverse group of proteins, including Runx2, p53, Smad3, CBP, and p300 (7-10), suggesting an important role for DDX5 in a multitude of developmental processes. Notably, DDX5 may be involved in growth factor-induced epithelial mesechymal transition (EMT). Phosphorylation of DDX5 at Tyr593 following PDGF stimulation was shown to displace Axin from β-catenin; this prevented phosphorylation of β-catenin by GSK-3β, leading to Wnt-independent nuclear translocation of β-catenin (11) and increased transcription of c-Myc, cyclin D1, and Snai1 (12,13).

  1. Ford, M.J. et al. (1988) Nature 332, 736-8.
  2. Hirling, H. et al. (1989) Nature 339, 562-4.
  3. Fukuda, T. et al. (2007) Nat Cell Biol 9, 604-11.
  4. Davis, B.N. et al. (2008) Nature 454, 56-61.
  5. Camats, M. et al. (2008) PLoS ONE 3, e2926.
  6. Wilson, B.J. et al. (2004) BMC Mol Biol 5, 11.
  7. Jensen, E.D. et al. (2008) J Cell Biochem 103, 1438-51.
  8. Bates, G.J. et al. (2005) EMBO J 24, 543-53.
  9. Warner, D.R. et al. (2004) Biochem Biophys Res Commun 324, 70-6.
  10. Rossow, K.L. and Janknecht, R. (2003) Oncogene 22, 151-6.
  11. Yang, L. et al. (2006) Cell 127, 139-55.
  12. Yang, L. et al. (2007) J Biol Chem 282, 16811-9.
  13. Carter, C.L. et al. (2010) Oncogene 29, 5427-36.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

For Research Use Only. Not For Use In Diagnostic Procedures.

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