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Product Pathways - Motif Antibodies

Phospho-(Ser/Thr) Akt Substrate Antibody #9611

Item# Description List Price Web Price Qty
9611L Phospho-(Ser/Thr) Akt Substrate Antibody - 300 µl $1,025.00
$922.50
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9611S Phospho-(Ser/Thr) Akt Substrate Antibody - 100 µl $433.00
$389.70
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9614S Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb - 100 µl $433.00
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9621L Phospho-(Ser/Thr) PKA Substrate Antibody - 300 µl $1,025.00
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9621S Phospho-(Ser/Thr) PKA Substrate Antibody - 100 µl $433.00
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9624S Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb - 100 µl $433.00
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity Source
W All Species Expected Endogenous Rabbit
IP
IHC-P
E-P

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), E-P=Peptide ELISA (DELFIA)

Specificity / Sensitivity

Phospho-(Ser/Thr) Akt Substrate Antibody preferentially recognizes peptides and proteins containing phospho-Ser/Thr preceded by Lys/Arg at positions -5 and -3, in a manner largely independent of other surrounding amino acids. Some cross-reactivity is observed for peptides that contain phospho-Ser/Thr preceded by Arg/Lys at positions -3 and -2. No cross-reactivity is observed with the corresponding nonphosphorylated sequences or with other phospho-Ser/Thr-containing motifs. By ELISA, the antibody recognizes a wide range of phosphorylated Akt substrate peptides, and, by 2-D gel Western blot analysis, it recognizes a large number of proteins presumed to be Akt substrates.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with phospho-Akt substrate peptides . Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or calyculin A-treated (0.1 µM for 30 minutes prior to lysis), using Phospho-(Ser/Thr) Akt Substrate Antibody. Proteins were separated by 2-D electrophoresis prior to blotting.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or calyculin A-treated, using Phospho-(Ser/Thr) Akt Substrate Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or insulin-treated (100 nM for 15 minutes prior to lysis), using Phospho-(Ser/Thr) Akt Substrate Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-(Ser/Thr) Akt Substrate Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-(Ser/Thr) Akt Substrate Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-(Ser/Thr) Akt Substrate Antibody.


ELISA-Peptide

ELISA-Peptide

Phospho-(Ser/Thr) Akt Substrate Antibody ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.)

Background

An important class of kinases, referred to as Arg-directed kinases or AGC-family kinases, includes cAMP-dependent protein kinase (PKA), cGMP-dependent protein kinase (PKG), protein kinase C, Akt, and RSK. These kinases share a substrate specificity characterized by Arg at position -3 relative to the phosphorylated Ser or Thr (1,2). Akt plays a central role in mediating critical cellular responses including cell growth and survival, angiogenesis, and transcriptional regulation (3-5). While a number of Akt substrates are known (such as GSK-3, Bad, and caspase-9) many important substrates await discovery. Akt phosphorylates substrates only at Ser/Thr in a conserved motif characterized by Arg at positions -5 and -3 (6). Phospho-Akt substrate-specific antibodies from Cell Signaling Technology are powerful tools for investigating the regulation of phosphorylation by Akt and other Arg-directed kinases, as well as for high throughput kinase drug discovery.

  1. Montminy, M. (1997) Annu Rev Biochem 66, 807-22.
  2. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  3. Marte, B.M. and Downward, J. (1997) Trends Biochem Sci 22, 355-8.
  4. Jiang, B.H. et al. (2000) Proc Natl Acad Sci USA 97, 1749-53.
  5. Scheid, M.P. and Woodgett, J.R. (2000) Curr Biol 10, R191-4.
  6. Alessi, D.R. et al. (1996) FEBS Lett 399, 333-8.

Application References

  • Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: Western Blotting.
  • Højlund, K. et al. (2008) Diabetes 57, 357-66. Applications: IP, Western Blotting.
  • Karlsson, H.K. et al. (2006) Diabetes 55, 1283-8. Applications: Western Blotting.
  • Srirangam, A. et al. (2006) Clin Cancer Res 12, 1883-96. Applications: IHC-P (paraffin).
  • Nascimento, E.B. et al. (2006) Diabetes 55, 3221-8. Applications: 2D Analysis, Western Blotting.
  • Trinidad, A.G. et al. (2006) J Leukoc Biol 79, 1073-82. Applications: Western Blotting.
  • Zhao, H. et al. (2005) J Neurosci 25, 9794-806. Applications: IF-P (paraffin).
  • Bruss, M.D. et al. (2005) Diabetes 54, 41-50. Applications: Western Blotting.
  • Wada, S. et al. (2005) J Biol Chem 280, 94-103. Applications: Western Blotting.
  • Burchfield, J.G. et al. (2004) J Biol Chem 279, 18623-32. Applications: Western Blotting.
  • Peng, X.D. et al. (2003) Genes Dev 17, 1352-65. Applications: Western Blotting.
  • Ly, C. et al. (2003) Cancer Res 63, 5716-22. Applications: 2D Analysis, IP, Western Blotting.
  • Kane, S. et al. (2002) J Biol Chem 277, 22115-8. Applications: IP, Western Blotting.
  • Lynch, D.K. and Daly, R.J. (2002) EMBO J 21, 72-82. Applications: Western Blotting.
  • Hermanson, O. et al. (2002) Nature 419, 934-9. Applications: Western Blotting.
  • Berwick, D.C. et al. (2002) J Biol Chem 277, 33895-900. Applications: Western Blotting.
  • Liu, M.Y. et al. (2002) Cancer Res 62, 6475-80. Applications: Western Blotting.
  • Wettey, F.R. et al. (2002) Science 297, 1521-5. Applications: Western Blotting.
  • Manning, B.D. et al. (2002) Mol Cell 10, 151-62. Applications: Western Blotting.
  • Fujita, N. et al. (2002) J Biol Chem 277, 28706-13. Applications: Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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