Revision 1

#30590Store at -20C

1 Kit

(4 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
PTEN (D4.3) XP® Rabbit mAb 9188 20 µl 54 kDa Rabbit IgG
Phospho-PTEN (Ser380/Thr382/383) (44A7) Rabbit mAb 9549 20 µl 54 kDa Rabbit IgG
PDK1 (D37A7) Rabbit mAb 5662 20 µl 58-68 kDa Rabbit IgG
Phospho-PDK1 (Ser241) (C49H2) Rabbit mAb 3438 20 µl 58 to 68 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The PTEN and PDK1 Antibody Sampler Kit II provides an economical means to evaluate two key enzymes that regulate multiple signaling pathways. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #30590

    PTEN and PDK1 Antibody Sampler Kit II

    PTEN and PDK1 Antibody Sampler Kit II: Image 1 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using PTEN (D4.3) XP® Rabbit mAb #9188. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    PTEN and PDK1 Antibody Sampler Kit II: Image 2 Expand Image
    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).
    PTEN and PDK1 Antibody Sampler Kit II: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using PDK1 (D37A7) Rabbit mAb.
    PTEN and PDK1 Antibody Sampler Kit II: Image 4 Expand Image
    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.
    PTEN and PDK1 Antibody Sampler Kit II: Image 5 Expand Image
    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).
    PTEN and PDK1 Antibody Sampler Kit II: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 7 Expand Image
    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.
    PTEN and PDK1 Antibody Sampler Kit II: Image 8 Expand Image
    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).
    PTEN and PDK1 Antibody Sampler Kit II: Image 9 Expand Image
    Western blot analysis of extracts from HCT 116 wild-type and HCT 116 PDK1-/- cells using PDK1 (D37A7) Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower). (HCT 116 wild-type and HCT 116 PDK1-/- cells were kindly provided by Dr. Bert Vogelstein, Johns Hopkins University, Baltimore, MD).
    PTEN and PDK1 Antibody Sampler Kit II: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using PTEN (D4.3) XP® Rabbit mAb performed on the Leica BOND RX.
    PTEN and PDK1 Antibody Sampler Kit II: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon using PTEN (D4.3) XP® Rabbit mAb.
    PTEN and PDK1 Antibody Sampler Kit II: Image 13 Expand Image
    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).