Cell Signaling Technology

Product Pathways - Akt Signaling

SignalSilence® PTEN siRNA (Human Specific) #6251

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with non-targeted (-) or targeted (+) siRNA. PTEN wasdetected using the PTEN Antibody #9552, and p42 was detected using the p42 MAPK Antibody #9108. The PTEN Antibody confirms silencing of PTEN expression, and the p42 MAPK Antibody is used to control for loading and specificity of PTEN siRNA.

Fluorescent Detection

Fluorescent Detection

Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.

Directions for Use

CST recommends transfection with 100 nM PTEN siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.

Background

PTEN (phosphatase and tensin homologue deleted on chromosome ten), also referred to as MMAC (mutated in multiple advanced cancers) phosphatase, is a tumor suppressor implicated in a wide variety of human cancers (1). PTEN encodes a 403 amino acid polypeptide originally described as a dual-specificity protein phosphatase (2). The main substrates of PTEN are inositol phospholipids generated by the activation of the phosphoinositide 3-kinase (PI3K) (3). PTEN is a major negative regulator of the PI3K/Akt signaling pathway (1,4,5). PTEN possesses a carboxy-terminal, noncatalytic regulatory domain with three phosphorylation sites (Ser380, Thr382 and Thr383) that regulate PTEN stability and may affect its biological activity (6,7). PTEN regulates p53 protein level and activity (8) and is involved in G protein coupled signaling during chemotaxis (9,10).

  1. Cantley, L.C. and Neel, B.G. (1999) Proc. Natl. Acad. Sci. USA 96, 4240-4245.
  2. Myers, M.P. et al. (1997) Proc. Natl. Acad. Sci. USA 94, 9052-9057.
  3. Myers, M.P. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 13513-13518.
  4. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-8211.
  5. Wu, X. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 15587-15591.
  6. Vazquez, F. et al. (2000) Mol. Cell. Biol. 20, 5010-5018.
  7. Torres, J. and Pulido, R. (2001) J. Biol. Chem. 276, 993-998.
  8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-130.
  9. Funamoto, S. et al. (2002) Cell 109, 611-623.
  10. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610.
  11. Czauderna, F. et al. (2003) Nucleic Acids Res. 31(2) , 670-682.

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.

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