Revision 3
#7978
Store at +4C
PathScan® Phospho-mTOR (Ser2481) Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H M
UniProt ID:
#P42345
Entrez-Gene Id:
#2475
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes | Product # | Quantity | Color | Storage Temp |
---|---|---|---|---|
mTOR Mouse mAb Coated Microwells | 76578 | 96 tests | +4C | |
Phospho-mTOR (Ser2481) Rabbit Detection mAb | 8468 | 1 ea | Green (Lyophilized) | +4C |
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) | 13272 | 1 ea | Red (Lyophilized) | +4C |
Detection Antibody Diluent | 13339 | 11 ml | Green | +4C |
HRP Diluent | 13515 | 11 ml | Red | +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 | Blue | +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
The PathScan® Phospho-mTOR (Ser2481) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of mTOR protein phosphorylated at Ser2481. A mTOR Mouse mAb has been coated onto the microwells. After incubation with cell lysates, mTOR (phospho and nonphospho) protein is captured by the coated antibody. Following extensive washing, Phospho-mTOR (Ser2481) Rabbit mAb is added to detect the captured phospho-mTOR protein. Anti-rabbit 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 mTOR phosphorylated at Ser2481.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
PathScan® Phospho-mTOR (Ser2481) Sandwich ELISA Kit #7978 detects endogenous levels of human mTOR protein when phosphorylated at Ser2481 as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Background
The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (1-3) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (4,5). When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1 (6). These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (7,8). mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy (9).
Background References
- Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
- Brown, E.J. et al. (1994) Nature 369, 756-8.
- Sabatini, D.M. et al. (1994) Cell 78, 35-43.
- Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
- Dennis, P.B. et al. (2001) Science 294, 1102-5.
- Fang, Y. et al. (2001) Science 294, 1942-5.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
- Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
- Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.
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 G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
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.
Revision 3
Figure 1. Treatment of HeLa cells with Calyculin A #9902 accumulates phosphorylation of mTOR at Ser2448 or Ser2481, as detected by PathScan® Phospho-mTOR (Ser2448) Sandwich ELISA Kit #7976 or PathScan® Phospho-mTOR (Ser2481) Sandwich ELISA Kit #7978, but does not affect levels of total mTOR protein detected by PathScan® Total mTOR Sandwich ELISA Kit #7974. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using mTOR (7C10) Rabbit mAb #2983, Phospho-mTOR (Ser2448) Antibody #2971 or Phospho-mTOR (Ser2481) Antibody #2974 is shown in the bottom figure.
Figure 2. The relationship between protein concentration of lysates from HeLa cells, untreated or reated with Calyculin A #9902, and the absorbance at 450 nm is shown. After starvation, HeLa cells (85% confluence) were treated with Calyculin A #9902 (10 nM) for 10-15 min at 37ºC and then lysed.