Product Pathways - PathScan ELISA
PathScan® Phospho-Akt (Thr308) Sandwich ELISA Antibody Pair #7144
|7144S||1 Kit (Reagents for 4 x 96 well plates)||---||In Stock||---|
|7144||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Volume||Cap Color|
|Akt Capture Ab (100X)||0.4 ml||Pink|
|P-Akt (T308) Detection Ab (100X)||0.4 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||0.04 ml||Yellow|
Storage: Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
CST's PathScan® Phospho-Akt (Thr308) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-Akt (Thr308) Sandwich ELISA Kit #7252. Capture and Detection Antibodies (100X stocks) and HRP-Conjugated Secondary Antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Akt Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by Phospho-Akt (Thr308) Mouse Detection Antibody and HRP-conjugated Anti-Mouse IgG. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Akt (Thr308) protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The relationship between lysate protein concentration from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-Akt (Thr308) Sandwich ELISA Antibody Pair #7144 is shown. After overnight starvation, NIH/3T3 cells were treated with PDGF (50 ng/ml) for 10 minutes at 37ºC and then lysed.
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).
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- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
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- 9803 Cell Lysis Buffer (10X)
- 9998 BSA
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.