Cell Signaling Technology

Product Pathways - Translational Control

mTOR Antibody #2972

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk (Pg) (Hr) Endogenous 289 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

mTOR Antibody detects endogenous levels of total mTOR. It cross-reacts weakly with some other proteins based on Western analysis.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2481 of human mTOR. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells (starved for 16 hours), untreated or EGF-treated (100 ng/ml), using Phospho-mTOR (Ser2448) antibody #2971 (upper) or mTOR Antibody (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with either control siRNA (-) or mTOR siRNA (+). mTOR was detected using mTOR Antibody #2972, and eIF4B was detected using eIF4B Antibody #3592. The mTOR Antibody confirms silencing of mTOR expression, and the eIF4B Antibody is used to control for loading and siRNA specificity.

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).

  1. Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
  2. Brown, E.J. et al. (1994) Nature 369, 756-8.
  3. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
  4. Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
  5. Dennis, P.B. et al. (2001) Science 294, 1102-5.
  6. Fang, Y. et al. (2001) Science 294, 1942-5.
  7. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
  8. Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
  9. Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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