| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Akt Rabbit mAb Coated Microwells | 56797 | 96 tests |
|
+4C |
| Akt2 Mouse Detection mAb | 13755 | 1 ea |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea |
|
+4C |
| Detection Antibody Diluent 2 | 14621 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
CST's PathScan® Total Akt2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Akt2 protein. An Akt rabbit antibody has been coated on the microwells. After incubation with cell lysates, Akt protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, Akt2 mouse antibody is added to detect captured Akt2 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 Akt2 protein.
*Antibodies in this kit are custom formulations specific to the kit.
Specificity/Sensitivity
Background
Akt, also referred to as PKB or Rac, plays a critical role in controlling cell 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 PI3K/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).
- Franke, T.F. et al. (1997) Cell 88, 435-7.
- Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
- Franke, T.F. et al. (1995) Cell 81, 727-36.
- Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
- Jacinto, E. et al. (2006) Cell 127, 125-37.
- Cardone, M.H. et al. (1998) Science 282, 1318-21.
- Brunet, A. et al. (1999) Cell 96, 857-68.
- Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
- Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
- Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
- Cross, D.A. et al. (1995) Nature 378, 785-9.
- Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
- Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
- Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
- Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
- Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
