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PDP - Template Name: FastScan ELISA Kit
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FastScan Phospho-Akt (Thr308) ELISA Kit #67807

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  • ELISA
ELISA Image 1: FastScan™ Phospho-Akt (Thr308) ELISA Kit
Figure 1. Treatment of NIH/3T3 cells with PDGF stimulates phosphorylation of Akt at Thr308 but does not affect the level of total Akt. The relationship between lysate protein concentration from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using the FastScan™ Phospho-Akt (Thr308) ELISA Kit #67807 is shown in the upper figure. The corresponding western blots using phospho-Akt (Thr308) antibody (left panel) and Akt antibody (right panel) are shown in the lower figure. After serum starvation, NIH/3T3 cells were treated with 100 ng/ml PDGF #8912 for 5 minutes at 37°C and then lysed.

To Purchase # 67807

Cat. # Size Qty. Price
67807C 1 Kit
96 assays
$654
67807V1 5 Kits
480 assays
$3,188
67807V2 10 Kits
960 assays
$6,213
67807V3 20 Kits
1920 assays
$12,099
67807V4 50 Kits
4800 assays
$29,430

Important Ordering Details

Custom Ordering Details:

If kit quantities from the same lot are needed in unlisted sizes, contact us for processing time and pricing.

Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.

Supporting Data

REACTIVITY H M R Mk
Application Key:
  • ELISA-ELISA 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • R-Rat 
  • Mk-Monkey 
  • Product Includes
  • Related Products
Product IncludesVolume (with Count)Solution Color
FastScan™ ELISA Microwell Strip Plate, 96 Well #532571 x 96 tests
Phospho-Akt (Thr308) Rabbit Capture mAb #850181 x 1 eaGreen (Lyophilized)
Akt (pan) Rabbit HRP-linked mAb #960471 x 1 eaRed (Lyophilized)
FastScan™ ELISA Capture Antibody Diluent #160761 x 3 mlGreen
FastScan™ ELISA HRP Antibody Diluent #281201 x 3 ml
TMB Substrate #70041 x 11 ml
STOP Solution #70021 x 11 ml
Sealing Tape #545031 x 1 ea
ELISA Wash Buffer (20X) #98011 x 25 ml
FastScan™ ELISA Cell Extraction Buffer (5X) #699051 x 10 ml
FastScan™ ELISA Cell Extraction Enhancer Solution (50X) #252431 x 1 ml
FastScan™ ELISA Kit #67807 Positive Control Type 2 #814361 x 1 ea

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Product Information

Product Description

The FastScan™ Phospho-Akt (Thr308) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Akt when phosphorylated at Thr308. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-Akt (Thr308) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-Akt (Thr308).

*Antibodies in this kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

The FastScan™ Phospho-Akt (Thr308) ELISA Kit detects endogenous levels of Akt when phosphorylated at Thr308 as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

Human, Mouse, Rat, Monkey

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).
  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.

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.
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