Cell Signaling Technology

Product Pathways - Translational Control

Phospho-PRAS40 (Thr246) Antibody #2640

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

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
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 (Thr246) Antibody detects endogenous levels of PRAS40 protein only when phosphorylated at Thr246.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the sequence surrounding Thr246 of human PRAS40. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, serum-starved for 14 hours and then either left untreated or treated with insulin (150 nM) for 15 minutes, using Phospho-PRAS40 (Thr246) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, serum-starved for 14 hours and then either left untreated or treated with insulin (150 nM) for 15 minutes, using Phospho-PRAS40 (Thr246) Antibody. The experiment was performed in the presence of phospho-peptide specific to phospho-PRAS40 (Thr246) (upper panel), the corresponding nonphospho-peptide (middle panel) and in the absence of any blocking peptide (lower panel).

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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products