Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

PTEN Blocking Peptide #1250

Description

This peptide is used to block PTEN (138G8) Rabbit mAb # 9559 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks PTEN (138G8) Rabbit mAb #9559 signal in immunohistochemistry.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 and HeLa cells using PTEN (138G6) Rabbit mAb # 9559 or the same antibody preincubated with PTEN Blocking Peptide.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using PTEN (138G8) Rabbit mAb #9559 preincubated with a control peptide (left) or with PTEN Blocking Peptide (right).

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. For western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 30 minutes before allowing to react with the blot. Recommended antibody dilutions can be found on the PTEN (138G6) Rabbit mAb #9559 data sheet.

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).

Applications: Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols and for western immunoblot analysis.Quality Control: The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks PTEN (138G8) Rabbit mAb # 9559 signal in immunohistochemistry.Directions for Use: For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the PTEN (138G6) Rabbit mAb #9559 data sheet.

  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

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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