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9652
PTEN and PDK1 Antibody Sampler Kit

PTEN and PDK1 Antibody Sampler Kit #9652

Western Blotting Image 1

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

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Western Blotting Image 2

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

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Western Blotting Image 3

Western blot analysis of extracts from PC3 cells, HCT116 wild-type and HCT116 PDK1 -/- cells using Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower). (HCT116 wild-type and HCT116 PDK1 -/- cells were kindly provided by Dr. Bert Vogelstein, Johns Hopkins University, Baltimore, MD).

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Western Blotting Image 4

Western blot analysis of extracts from SW-13, NIH/3T3, Jurkat and PC12 cells, using PDK1 Antibody.

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Western Blotting Image 5

Western blot analysis of extracts from various cell lines using Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) Rabbit mAb. 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.

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Western Blotting Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IHC-P (paraffin) Image 7

Immunohistochemical analysis of paraffin-embedded human colon using PTEN (D4.3) XP® Rabbit mAb.

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Western Blotting Image 8

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

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IHC-P (paraffin) Image 9

Immunohistochemical analysis using PTEN (D4.3) XP® Rabbit mAb on SignalSlide(TM) PTEN IHC Controls #8106 (paraffin-embedded LNCaP (left) and NIH/3T3 (right) cells).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb 9549 20 µl
  • WB
H M R Mk 54 Rabbit IgG
PTEN (D4.3) XP® Rabbit mAb 9188 20 µl
  • WB
  • IP
  • IHC
H M R Mk Dg 54 Rabbit IgG
Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb 3438 20 µl
  • WB
  • IP
H M R 58 to 68 Rabbit IgG
PDK1 Antibody 3062 20 µl
  • WB
H M R Mk 58 to 68 Rabbit 
Non-phospho PTEN (Ser380/Thr382/Thr383) (D2D11) Rabbit mAb 7960 20 µl
  • WB
  • IP
H M R Mk 55 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two Western blots per primary antibody.

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.

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.

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 LJ (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-R96.
  12. Toker, A. and Newton, A.C. (2000) Cell 103, 185-188.
  13. Williams, M.R. et al. (2000) Curr. Biol. 10, 439-448.
Entrez-Gene Id
5170 , 5728
Swiss-Prot Acc.
O15530 , P60484
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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