Product Pathways - PathScan ELISA
PathScan® Phospho-Akt (Thr308) Chemiluminescent Sandwich ELISA Kit #7135
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|Kit Includes||Volume||Solution Color|
|Akt Rabbit mAb Coated Microwells||96 tests|
|Phospho-Akt (Thr308) Mouse Detection mAb||5.5 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Phospho-Akt (Thr308) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Akt (Thr308) protein with a chemiluminescent readout. Chemiluminescence ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. An Akt rabbit antibody has been coated on the microwells. After incubation with cell lysates, both phospho- and nonphospho-Akt proteins are captured by the coated antibody. Following extensive washing, phospho-Akt (Thr308) mouse antibody is added to detect the captured phospho-Akt protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-Akt (Thr308) protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-Akt (Thr308) Chemiluminescent Sandwich ELISA Kit #7135 detects endogenous levels of phospho-Akt (Thr308) in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Relationship between protein concentration of lysates from untreated and PDGF-treated NIH/3T3 cells and immediate light generation with chemiluminescent substrate is shown. Cells (80% confluence) were treated with PDGF #9909 (50 ng/ml) and lysed after incubation at 37ºC for 5 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
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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* Product-specific protocol.
- 7252 PathScan® Phospho-Akt (Thr308) Sandwich ELISA Kit
- 7144 PathScan® Phospho-Akt (Thr308) Sandwich ELISA Antibody Pair
- 7170 PathScan® Total Akt1 Sandwich ELISA Kit
- 7142 PathScan® Total Akt1 Sandwich ELISA Antibody Pair
- 7160 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
- 7143 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Antibody Pair
- 4685 Akt (pan) (11E7) Rabbit mAb
- 4691 Akt (pan) (C67E7) Rabbit mAb
- 2920 Akt (pan) (40D4) Mouse mAb
- 9272 Akt Antibody
- 2938 Akt1 (C73H10) Rabbit mAb
- 2967 Akt1 (2H10) Mouse mAb
- 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
- 4056 Phospho-Akt (Thr308) (244F9) Rabbit mAb
- 5106 Phospho-Akt (Thr308) (L32A4) Mouse mAb
- 9275 Phospho-Akt (Thr308) Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.