Cell Signaling Technology

Product Pathways - Translational Control

Phospho-PRAS40 (Ser183) Antibody #5936

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M (R) Endogenous 40 Rabbit

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

Protocols

Specificity / Sensitivity

Phospho-PRAS40 (Ser183) Antibody recognizes endogenous levels of PRAS40 protein only when phosphorylated at Ser183.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser183 of human PRAS40 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with LY294002 #9901 (with and without serum starvation) or with Rapamycin #9904, using Phospho-PRAS40 (Ser183) Antibody (upper) or PRAS40 (D23C7) XP® Rabbit mAb #2691 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated with hIGF-I #8917, in the absence (-) or presence (+) of λ phosphatase, using Phospho-PRAS40 (Ser183) Antibody (upper) or PRAS40 (D23C7) XP® Rabbit mAb #2691 (lower).

Background

Many growth factors and hormones induce the phosphoinositide 3-kinase signaling pathway, which results in the activation of downstream effector proteins such as the serine/threonine kinase Akt (1,2). One known Akt substrate is a 40 kDa, proline-rich protein (PRAS40) that binds to 14-3-3 proteins (2). PRAS40 also binds mTOR to transduce Akt signals to the mTOR complex. Inhibition of mTOR signaling stimulates PRAS40 binding to mTOR, which in turn inhibits mTOR activity (3). PRAS40 interacts with raptor in mTOR complex 1 (mTORC1) in insulin-deprived cells and inhibits the activation of the mTORC1 pathway mediated by the cell cycle protein Rheb. Phosphorylation of PRAS40 by Akt at Thr246 relieves PRAS40 inhibition of mTORC1 (4). mTORC1 in turn phosphorylates PRAS40 at Ser183 (5).

  1. Cantley, L.C. (2002) Science 296, 1655-7.
  2. Kovacina, K.S. et al. (2003) J Biol Chem 278, 10189-94.
  3. Vander Haar, E. et al. (2007) Nat Cell Biol 9, 316-23.
  4. Sancak, Y. et al. (2007) Mol Cell 25, 903-15.
  5. Oshiro, N. et al. (2007) J Biol Chem 282, 20329-39.

Application References

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