Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

PTEN and PDK1 Antibody Sampler Kit #9652

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb #9549 40 µl W H M R Mk 54 Rabbit IgG
PTEN (D4.3) XP® Rabbit mAb #9188 40 µl W IP IHC-P H M R Mk Dg (C) 54 Rabbit IgG
Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb #3438 40 µl W IP H M R 58 to 68 Rabbit IgG
PDK1 Antibody #3062 40 µl W H M R Mk 58 to 68 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) Rabbit mAb #7960 40 µl W IP H M R Mk 55 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the PTEN and PDK1 Antibody Sampler Kit detects endogenous levels of its target protein. Activation state antibodies detect only target proteins phosphorylated at indicated residues. Non-Phospho-PTEN (Ser380/Thr382/Thr383) Antibody detects endogenous levels of PTEN only when dephosphorylated at Ser380, Thr382 and Thr383.

Western Blotting

Western Blotting

Western blot analysis of extracts from SW-13, NIH/3T3, Jurkat and PC-12 cells using PDK1 Antibody #3062.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC3 cells, untreated or λ phosphatase-treated, using Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb #3438 (upper), PDK1 Antibody #3062 (middle) or Akt Antibody #9272 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) Rabbit mAb #7960. The non-phosphospecificity of the antibody was verified by preincubating the antibody without peptide (-), with PTEN (Ser380/Thr382/Thr383) non-phosphopeptide (+), or with PTEN (Ser380/Thr382/Thr383) phosphopeptide (+) prior to incubating the membrane.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PTEN (D4.3) XP® Rabbit mAb #9188 (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells using Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb #9549. Membranes were left untreated (-) or treated with (+) calf intestinal phosphatase (CIP) following western transfer to verify phospho-specificity of the antibody.

Description

The PTEN and PDK1 Sampler Kit provides an economical means to evaluate two key enzymes that regulate multiple signaling pathways. The kit contains enough primary and secondary antibodies to perform four Western blots per primary antibody.

Source / Purification

Phospho-specific rabbit monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues around Ser380, Thr382 and Thr383 of human PTEN and around Ser241 of human PDK1. PTEN (D4.3) XP® Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of human PTEN. Monoclonal antibody Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) is produced by immunizing animals with a synthetic non-phosphopeptide corresponding to residues surrounding Ser380/Thr382/Thr383 of human PTEN protein. Polyclonal PDK1 antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding the carboxy terminus of human PDK1. Polyclonal antibodies are purified using protein A and peptide affinity chromatography.

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).

Phosphoinositide-dependent protein kinase 1 (PDK1) plays a central role in many signal transduction pathways (11,12) including the activation of Akt and the PKC isoenzymes p70 S6 kinase and RSK (13). Through its effects on these kinases, PDK1 is involved in the regulation of a wide variety of processes, including cell proliferation, differentiation and apoptosis.

  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.
  11. Belham, C. et al. (1999) Curr Biol 9, R93-6.
  12. Toker, A. and Newton, A.C. (2000) Cell 103, 185-8.
  13. Williams, M.R. et al. (2000) Curr Biol 10, 439-48.

Application References

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Protocols

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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