Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

SignalSilence® γ-Catenin siRNA I #6226

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® γ-Catenin siRNA I (+) or SignalSilence® γ-Catenin siRNA II #6239 (+), 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 γ-catenin expression silencing. The α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® γ-Catenin siRNA I (+) or SignalSilence® γ-Catenin siRNA II #6239 (+), using γ-Catenin Antibody #2309 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The γ-Catenin Antibody confirms silencing of γ-catenin expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Description

SignalSilence® γ-Catenin siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit γ-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.

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 γ-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.

Background

Also known as plakoglobin, γ-catenin is a member of the Armadillo family of signaling molecules, which includes β-catenin and the Drosophila protein armadillo (1). This family of proteins is involved in Wnt signaling, which is important in embryonic development and in tumorigenesis (2-3). Although the two vertebrate proteins β- and γ-catenin display sequence homology, γ-catenin likely plays a role distinct from that of β-catenin (1, 4-6). γ-catenin localizes to desmosomes and adherens junctions, both sites of intercellular adhesion, and interacts with the cytoplasmic domains of classical and desmosomal cadherins. Interaction of γ- or β-catenin with α-catenin, desmoplakin and other junction proteins provides a link between intercellular junctions and the actin and intermediate filament cytoskeleton. Maintenance and/or modification of this link is vital for control of cell adhesion and migration (1). γ-catenin is modified by phosphorylation, affecting both adhesion and β-catenin dependent transcription (7), and by and O-glycosylation, affecting adhesion (8). Recent evidence suggests that γ-catenin regulates desmosomal adhesion in response to growth factor stimulation (9).

  1. Zhurinsky, J. et al. (2000) J. Cell Sci. 113 ( Pt 18), 3127-3139.
  2. Wodarz, A. and Nusse, R. (1998) Annu. Rev. Cell Dev. Biol. 14, 59-88.
  3. Polakis, P. (1999) Curr. Opin. Genet. Dev. 9, 15-21.
  4. Zhurinsky, J. et al. (2000) Mol. Cell Biol. 20, 4238-4252.
  5. Charpentier, E. et al. (2000) J. Cell Biol. 149, 503-520.
  6. Kolligs, F.T. et al. (2000) Genes Dev. 14, 1319-13131.
  7. Miravet, S. et al. (2003) Mol. Cell Biol. 23, 7391-7402.
  8. Hu, P. et al. (2006) J. Biol. Chem. 281, 12786-12791.
  9. Yin, T. et al. (2005) J. Biol. Chem. 280, 40355-40363.

Application References

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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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