Product Pathways - Jak/Stat Pathway
SignalSilence® Stat6 siRNA II #9404
|Transfection||H M (R) (Mk)|
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Western blot analysis of extracts from L-929 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Stat6 siRNA I (Mouse Specific) #9396 (+), or SignalSilence® Stat6 siRNA II (+), using Stat6 (D3H4) Rabbit mAb #5397 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Stat6 (D3H4) Rabbit mAb confirms silencing of Stat6 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
SignalSilence® Stat6 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Stat6 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.
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 SignalSilence® Stat6 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.
Upon activation by Janus kinases, Stat6 translocates to the nucleus where it regulates cytokine-induced gene expression. Stat6 is activated via phosphorylation at Tyr641 and is required for responsiveness to IL-4 and IL-13 (1-4). In addition, Stat6 is activated by IFN-α in B cells, where it forms transcriptionally active complexes with Stat2 and p48 (5,6). Protein phosphatase 2A is also involved in regulation of IL-4-mediated Stat6 signaling (7).
- Nelms, K. et al. (1999) Ann. Rev. Immunol. 17, 701-738.
- Malabarba, M.G. et al. (1996) Biochem. J. 319, 865-872.
- Hou, J. et al. (1994) Science 265, 1701-1706.
- Quelle, F.W. et al. (1995) Mol. Cell. Biol. 15, 3336-3343.
- Takeda, K. et al. (1996) Nature 380, 627-630.
- Gupta, S. et al. (1999) J. Immunol. 163, 3834-3841.
- Woetmann, A. et al. (2003) J. Biol. Chem. 278, 2787-2791.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 9396 SignalSilence® Stat6 siRNA I (Mouse Specific)
- 5397 Stat6 (D3H4) Rabbit mAb
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.