Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

SignalSilence® PTEN siRNA II #6538

Applications Reactivity
Transfection H (M) (R) (Mk)

Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
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 (Fluorescein Conjugate) #6201 (-) or SignalSilence® PTEN siRNA II (+), using PTEN (138G6) Rabbit mAb #9559 and β-Actin (13E5) Rabbit mAb #4970. PTEN (138G6) Rabbit mAb confirms silencing of PTEN expression and β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of PTEN siRNA.

Description

SignalSilence® PTEN siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PTEN 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 are rigorously tested in-house and have been shown to reduce 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 PTEN 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.

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 levels 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, 4240-5.
  2. Myers, M.P. et al. (1997) Proc Natl Acad Sci USA 94, 9052-7.
  3. Myers, M.P. et al. (1998) Proc Natl Acad Sci USA 95, 13513-8.
  4. Wan, X. and Helman, L.J. (2003) Oncogene 22, 8205-11.
  5. Wu, X. et al. (1998) Proc Natl Acad Sci USA 95, 15587-91.
  6. Vazquez, F. et al. (2000) Mol Cell Biol 20, 5010-8.
  7. Torres, J. and Pulido, R. (2001) J Biol Chem 276, 993-8.
  8. Freeman, D.J. et al. (2003) Cancer Cell 3, 117-30.
  9. Funamoto, S. et al. (2002) Cell 109, 611-23.
  10. Iijima, M. and Devreotes, P. (2002) Cell 109, 599-610.

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.

Products