Product Pathways - Translational Control
Phospho-4E-BP1 (Ser65) Antibody #9451
|W IP||H M R Mk||Endogenous||15 to 20||Rabbit|
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.
Specificity / Sensitivity
Phospho-4E-BP1 (Ser65) Antibody detects endogenous levels of 4E-BP1 when phosphorylated at serine 65. This antibody has been shown to also recognize 4E-BP1 when phosphorylated at serine 101 (See application reference, Wang et al. 2003).
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the sequence of mouse 4E-BP1. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from 293 cells using 4E-BP1 Antibody #9644 (lower) and Phospho-4E-BP1 (Ser65) Antibody #9451 (upper). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes.
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).
- Pause, A. et al. (1994) Nature 371, 762-767.
- Brunn, G.J. et al. (1997) Science 277, 99-101.
- Gingras, A.C. et al. (1998) Genes Dev. 12, 502-513.
- Fadden, P. et al. (1997) J. Biol. Chem. 272, 10240-10247.
- Gingras, A.C. et al. (1999) Genes Dev. 13, 1422-1437.
- Aoki, M. et al. (2001) A role of the kinase mTOR in cellular transformation induced by the oncoproteins P3k and Akt. Proc. Natl. Acad. Sci. USA 98, 136-141. Applications: Western Blotting
- Jiang, Y. P. et al. (2001) Rapamycin-insensitive regulation of 4e-bp1 in regenerating rat liver. J. Biol. Chem. 276, 10943-10951. Applications: Western Blotting
- Wang, X. et al. (2003) The C Terminus of Initiation Factor 4E-Binding Protein 1 Contains Multiple Regulatory Features That Influence Its Function and Phosphorylation. Mol. Cell. Biol. 23, 1546-1557. Applications: Western Blotting
- Gingras, A. C. et al. (2001) Hierarchical phosphorylation of the translation inhibitor 4E-BP1. Genes Dev. 15, 2852-2864. Applications: 2D Gel Electrophoresis
- Ma, D. et al. (2009) J Cell Physiol 219, 31-44. Applications: Western Blotting
- Fonseca, B.D. et al. (2011) J Biol Chem 286, 27111-22. Applications: IP
- Ito, N. et al. (2011) Lab Invest 91, 1584-95. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.