Upstream / Downstream

Explore pathways related to this product.

Antibody Guarantee

CST Antibody Performance Guarantee

LEARN MORE  

To Purchase # 1140S

1140S 100 µg $79.00
$ 0. 00

Questions?

Find answers on our FAQs page.

ANSWERS  

Visit PhosphoSitePlus®

PTM information and tools available.

LEARN MORE

Immunohistochemical analysis of paraffin-embedded SK-OV3 metastatic tumor in mouse lung using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 in the presence of control peptide (left) or Phospho-Akt (Ser473) Blocking Peptide (right).

Learn more about how we got this image

Flow cytometric analysis of Jurkat cells, untreated (blue) or LY294002 and Wortmannin-treated (red), using Phospho-Akt (Ser473) (193H12) Rabbit mAb #4058. The treatment-induced shift in fluorescence (left) was abolished when phospho-Akt antibody was preincubated with Phospho-Akt (Ser473) Blocking Peptide #1140 (right).

Learn more about how we got this image
Image

Product Description

This peptide is used to block Phospho-Akt (Ser473) (193H12) Rabbit mAb #4058, Phospho-Akt (Ser473) (736E11) Rabbit mAb (IHC Specific) #3787 and Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 reactivity.


Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-Akt (Ser473) Rabbit mAb #4060 and (IHC Specific) #3787 signal completely in immunohistochemistry and Phospho-Akt (Ser473) (193H12) Rabbit mAb #4058 signal in flow cytometry.

Product Usage Information

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and flow cytometry protocols. For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. For flow cytometry, add antibody at the recommended dilution to 100 µl incubation buffer. Next, add 10 µl blocking peptide. Incubate at room temperature for 15-30 minutes before applying to cells. *Recommended antibody dilutions can be found on the antibody datasheet.


Storage: Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA and 5% glycerol. Store at –20°C.

Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).


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

2.  Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.

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

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

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

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

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

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

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

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

11.  Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.

12.  Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.

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

14.  Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.

15.  Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.

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

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

18.  Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.

19.  Manning, B.D. et al. (2002) Mol Cell 10, 151-62.


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


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.