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Custom Ordering Details: The control slides that accompany this kit are cut freshly upon ordering. Please allow up to three business days for your product to be processed.

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Schematic of placement of cell pellets on SignalSlide® Akt Family IHC Controls #8115.

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Product Includes

Kit Includes Quantity Antigen Retrieval / Diluent Isotype
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 40 µl Citrate / SignalStain® Antibody Diluent #8112 Rabbit IgG
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb #4858 40 µl Citrate / SignalStain® Antibody Diluent #8112 Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb #4691 40 µl Citrate / SignalStain® Antibody Diluent #8112 Rabbit IgG
PTEN (D4.3) XP® Rabbit mAb #9188 40 µl Citrate / SignalStain® Antibody Diluent #8112 Rabbit IgG
*SignalStain® Antibody Diluent #8112 25 ml
SignalSlide® Akt Family IHC Controls #8115 1 Pack
*SignalStain® Antibody Diluent is supplied as a working solution and should be stored at 4ºC (packaged separately).
†Control slides should be stored at 4ºC (packaged separately).

Product Description

The SignalStain® Akt Pathway IHC Sampler Kit from Cell Signaling Technology allows the researcher to examine paraffin-embedded tissues or cells with antibodies directed against proteins involved in Akt signaling. Multiple approaches are used to validate each antibody for use in immunohistochemical assays. Also included in the kit are control slides that can be used to verify the performance of each antibody and a primary antibody diluent. Please see the above table for the recommended antibody diluent for each kit antibody.


Specificity / Sensitivity

Each antibody in the SignalStain® Akt Pathway IHC Sampler Kit detects endogenous levels of its target protein and does not cross-react with related proteins.


Source / Purification

Monoclonal antibody is produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding the phosphorylation site(s) of interest, and synthetic peptides corresponding to the carboxy-terminal sequence of mouse Akt or from the carboxy-terminal sequence of human PTEN.

The protein kinase Akt (also called PKB or Rac) is activated by insulin and growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (1,2). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (3) and by carboxy terminal phosphorylation at Ser473 by the mTORC2 complex composed of mammalian target of rapamycin (mTOR) in a complex with rictor and Sin1 (4,5). Akt signaling is negatively regulated via the PTEN phosphatase (6).

Akt’s several functions include inhibition of apoptosis (7-9), regulation of glycogen synthesis via GSK-3 (10), and promotion of the cell cycle (11). Akt also regulates protein synthesis by phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (mTORC1) (12). Akt also effects mTOR activity via phosphorylation and inhibition of PRAS40 (40 kDa, proline-rich protein), which binds to raptor in the mTORC1 complex and inhibits mTOR activity (13). Phosphorylation of PRAS40 by Akt at Thr246 relieves PRAS40 inhibition of mTORC1 (14), allowing protein synthesis to occur. Active of mTORC1 signals to p70 S6 kinase, which in turn phosphorylates S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation of a subset of mRNA transcripts that encode ribosomal proteins, translation elongation factors as well as regulators of cell cycle progression (15). Important S6 ribosomal protein phosphorylation sites include Ser235, Ser236, Ser240 and Ser244 within a small carboxy-terminal region (16).


1.  Franke, T.F. et al. (1997) Cell 88, 435-7.

2.  Peterson, R.T. and Schreiber, S.L. (1998) Curr Biol 8, R248-50.

3.  Franke, T.F. et al. (1995) Cell 81, 727-36.

4.  Cross, D.A. et al. (1995) Nature 378, 785-9.

5.  Vander Haar, E. et al. (2007) Nat Cell Biol 9, 316-23.

6.  Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.

7.  Sancak, Y. et al. (2007) Mol Cell 25, 903-15.

8.  Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5.

9.  Sarbassov, D.D. et al. (2005) Science 307, 1098-101.

10.  Jacinto, E. et al. (2006) Cell 127, 125-37.

11.  Cardone, M.H. et al. (1998) Science 282, 1318-21.

12.  Brunet, A. et al. (1999) Cell 96, 857-68.

13.  Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.

14.  Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.

15.  Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.

16.  Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.


Entrez-Gene Id 207, 208, 10000
Swiss-Prot Acc. P31749, P31751, Q9Y243

Protein Specific References

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Hill MM et al. (2001) J Biol Chem 276, 25643–6

Dhawan P et al. (2002) Cancer Res 62, 7335–42

Conus NM et al. (2002) J Biol Chem 277, 38021–8

Sano H et al. (2002) J Biol Chem 277, 19439–47

Egawa K et al. (2002) J Biol Chem 277, 38863–9

Kisseleva MV et al. (2002) J Biol Chem 277, 6266–72

Barry FA and Gibbins JM (2002) J Biol Chem 277, 12874–8

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Rani MR et al. (2002) J Biol Chem 277, 38456–61

Ho R et al. (2002) Cancer Res 62, 6462–6

Wan X and Helman LJ (2003) Oncogene 22, 8205–11

Fukuda T et al. (2003) J Biol Chem 278, 51324–33

Kim HH et al. (2003) FASEB J 17, 2163–5

Min YH et al. (2004) Cancer Res 64, 5225–31

Tazzari PL et al. (2004) Br J Haematol 126, 675–81

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Wolfrum S et al. (2004) Arterioscler Thromb Vasc Biol 24, 1842–7

Kaneko Y et al. (2004) J Cell Sci 117, 407–15

Esfandiarei M et al. (2004) J Virol 78, 4289–98

Baudhuin LM et al. (2004) FASEB J 18, 341–3

Dietze EC et al. (2004) Oncogene 23, 3851–62

Wu T et al. (2004) Mol Cancer Ther 3, 299–307

Honjo S et al. (2005) DNA Cell Biol 24, 141–7

Karlsson HK et al. (2005) Diabetes 54, 1459–67

Viniegra JG et al. (2005) J Biol Chem 280, 4029–36

Le XF et al. (2005) J Biol Chem 280, 2092–104

Smith E and Frenkel B (2005) J Biol Chem 280, 2388–94

Edwards LA et al. (2005) Oncogene 24, 3596–605

Karlsson HK et al. (2005) Diabetes 54, 1692–7

Kippenberger S et al. (2005) J Biol Chem 280, 3060–7

Jung HS et al. (2005) Mol Endocrinol 19, 2748–59

Khundmiri SJ et al. (2006) Am J Physiol Cell Physiol 291, C1247–57

Hers I and (2007) Blood 110, 4243–52

Ananthanarayanan B et al. (2007) J Biol Chem 282, 36634–41

Zunder ER et al. (2008) Cancer Cell 14, 180–92

Grenegård M et al. (2008) J Biol Chem 283, 18493–504

Abubaker J et al. (2009) Mol Cancer 8, 51

Chen PL and Easton AS (2011) Curr Neurovasc Res 8, 14–24

Van Aller GS et al. (2011) Biochem Biophys Res Commun 406, 194–9

Uesugi A et al. (2011) Cancer Res 71, 5765–78

Ou YH et al. (2011) Mol Cell 41, 458–70

Wang S et al. (2012) PLoS One 7, e37427

Glidden EJ et al. (2012) J Biol Chem 287, 581–8

Shih MC et al. (2012) Oncogene 31, 2389–400

Misra UK and Pizzo SV (2012) J Cell Biochem 113, 1488–500

Johnson AL et al. (2001) Biol Reprod 64, 1566–74

Zhang M and Riedel H (2009) J Cell Biochem 107, 65–75


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
SignalStain® is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 5,675,063.