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REACTIVITY
Product Includes Volume Solution Color
Akt Rabbit mAb Coated Microwells 96 tests
Akt3 (L36F12) Mouse Detection mAb 1 Ea Green (Lyophilized)
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 1 Ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

The PathScan® Total Akt3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Akt3 protein. An Akt rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Akt protein is captured by the coated antibody. Following extensive washing, an Akt3 mouse mAb is added to detect captured Akt3 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Akt3 protein.

Antibodies in kit are custom formulations specific to kit.


Specificity / Sensitivity

PathScan® Total Akt3 Sandwich ELISA Kit (Mouse Preferred) detects endogenous levels of Akt3 protein in mouse cells, as shown in Figure 1. Akt3 kit specificity is demonstrated in Figure 2, while the kit sensitivity is shown in Figure 3. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.


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

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

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

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

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

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

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

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

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

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

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

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 10000
Swiss-Prot Acc. Q9Y243


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