Cell Signaling Technology

Product Pathways - Translational Control

Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb #4923

Applications Reactivity Sensitivity MW (kDa) Isotype
W F H M R Mk Endogenous 15-20 Rabbit IgG

Applications Key:  W=Western Blotting  F=Flow Cytometry
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

Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb detects endogenous levels of 4E-BP1 only when dephosphorylated at Thr46. The antibody cross-reacts with 4E-BP2 and 4E-BP3 dephosphorylated at equivalent sites.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr46 of human 4E-BP1.  

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, treated with λ phosphatase or 20% FBS as indicated, using Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb (upper), Phospho-4E-BP1 (Thr37/46) Antibody #9459 (middle), and 4E-BP1 Antibody #9452 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of COS cells, untreated (blue) or λ phosphatase-treated (green), using Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb compared to a nonspecific negative control antibody (red).

Background

Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR at Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).

  1. Pause, A. et al. (1994) Nature 371, 762-767.
  2. Brunn, G.J. et al. (1997) Science 277, 99-101.
  3. Gingras, A.C. et al. (1998) Genes Dev. 12, 502-513.
  4. Fadden, P. et al. (1997) J. Biol. Chem. 272, 10240-10247.
  5. Gingras, A.C. et al. (1999) Genes Dev. 13, 1422-1437.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.


This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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