Product Pathways - Development
SignalSilence® β-Catenin siRNA I #6225
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 HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® β-Catenin siRNA I (+) or SignalSilence® β-Catenin siRNA II #6238 (+), using β-Catenin (6B3) Rabbit mAb #9582 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The β-catenin antibody confirms silencing of β-catenin expression, while the α-tubulin rabbit mAb is used is used as a loading control.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® β-Catenin siRNA I (+) or SignalSilence® β-Catenin siRNA II #6238 (+), using β-Catenin Antibody #9562 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The β-catenin antibody recognizes both β-catenin and γ-catenin and demonstrates the specificity of β-catenin expression silencing. The α-tubulin rabbit mAb is used is used as a loading control.
SignalSilence® β-Catenin siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit β-catenin 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 β-Catenin 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.
β-Catenin is a key downstream effector in the Wnt signaling pathway (1). It is implicated in two major biological processes in vertebrates: early embryonic development (2) and tumorigenesis (3). CK1 phosphorylates β-catenin at Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3β (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37, and Thr41 (7). Mutations at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines (8).
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- Wodarz, A. and Nusse, R. (1998) Annu. Rev. Cell. Dev. Biol. 14, 59-88.
- Polakis, P. (1999) Curr. Opin. Genet. Dev. 9, 15-21.
- Amit, S. et al. (2002) Genes Dev. 16, 1066-1076.
- Lin, C. et al. (2002) Cell 108, 837-847.
- Yanagawa, S. et al. (2002) EMBO J. 21, 1733-1742.
- Yost, C. et al. (1996) Genes Dev. 10, 1443-1454.
- Morin, P.J. (1997) Science 275, 1787-1790.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 6238 SignalSilence® β-Catenin siRNA II
- 9582 β-Catenin (6B3) Rabbit mAb
- 9562 β-Catenin Antibody
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.