Cell Signaling Technology

Product Pathways - MAPK Signaling

SignalSilence® c-Jun siRNA I #6203

Applications Reactivity
Transfection H

Reactivity Key:  H=Human
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® c-Jun siRNA I (+) or SignalSilence® c-Jun siRNA II #6204 (+), using c-Jun (60A8) Rabbit mAb #9165 and α-Tubulin (11H10) Rabbit mAb #2125. The c-Jun (60A8) Rabbit mAb confirms silencing of c-Jun expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of c-Jun siRNA.

Description

SignalSilence® c-Jun siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit c-Jun 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 c-Jun siRNA I 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

c-Jun is a member of the Jun family containing c-Jun, JunB, and JunD, and is a component of the transcription factor activator protein-1 (AP-1). AP-1 is composed of dimers of Fos, Jun, and ATF family members and binds to and activates transcription at TRE/AP-1 elements (reviewed in 1). Extracellular signals including growth factors, chemokines, and stress activate AP-1-dependent transcription. The transcriptional activity of c-Jun is regulated by phosphorylation at Ser63 and Ser73 through SAPK/JNK (reviewed in 2). Knock-out studies in mice have shown that c-Jun is essential for embryogenesis (3), and subsequent studies have demonstrated roles for c-Jun in various tissues and developmental processes including axon regeneration (4), liver regeneration (5), and T cell development (6). AP-1 regulated genes exert diverse biological functions including cell proliferation, differentiation, and apoptosis, as well as transformation, invasion and metastasis, depending on cell type and context (7-9). Other target genes regulate survival, as well as hypoxia and angiogenesis (8,10). Research studies have implicated c-Jun as a promising therapeutic target for cancer, vascular remodeling, acute inflammation, and rheumatoid arthritis (11,12).

  1. Jochum, W. et al. (2001) Oncogene 20, 2401-12.
  2. Davis, R.J. (2000) Cell 103, 239-52.
  3. Hilberg, F. et al. (1993) Nature 365, 179-81.
  4. Raivich, G. et al. (2004) Neuron 43, 57-67.
  5. Behrens, A. et al. (2002) EMBO J 21, 1782-90.
  6. Riera-Sans, L. and Behrens, A. (2007) J Immunol 178, 5690-700.
  7. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  8. Shaulian, E. and Karin, M. (2002) Nat Cell Biol 4, E131-6.
  9. Weiss, C. and Bohmann, D. (2004) Cell Cycle 3, 111-3.
  10. Karamouzis, M.V. et al. (2007) Mol Cancer Res 5, 109-20.
  11. Kim, S. and Iwao, H. (2003) J Pharmacol Sci 91, 177-81.
  12. Dass, C.R. and Choong, P.F. (2008) Pharmazie 63, 411-4.

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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