Technical Data
4E Binding Protein 1, phosphorylated (Thr37/46) (Protein Synthesis Initiation Factor 4E Binding Protein, eIF-4EBP1, PHAS-1, PHAS-I, Eukaryotic Translation Initiation Factor BP)
When bound to eIF4E, 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation. Upon hyperphosphorylation of 4E-BP1 this interaction is disrupted and cap-dependent translation is activated (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 on 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).

Suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence/Immunocytochemistry, and Flow cytometry. Other applications have not been tested.

Recommended Dilutions:
Western Blot: 1:1000. For Western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween-20 at 4C with gentle shaking, overnight.
Immunohistochemistry (Paraffin): 1:1600 (Citrate/TBST)
Immunofluorescence (IC): 1:200
Flow Cytometry: 1:50
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
100ul-20CBlue IceMouseRabbit
Not determined
Synthetic phospho-peptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1, conjugated to KLH.
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, <0.02% sodium azide, 50% glycerol.
Recognizes endogenous levels of mouse 4E Binding Protein 1 only when phosphorylated at Thr37 and/or Thr46. May crossreact with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites. Species Crossreactivity: human, rat, monkey, and D. melanogaster.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Pause, A. et al. (1994) Nature 371, 762767. (2) Brunn, G.J. et al. (1997) Science 277, 99101.
(3) Gingras, A.C. et al. (1998) Genes Dev. 12, 502513. (4) Fadden, P. et al. (1997) J. Biol. Chem. 272, 1024010247. (5) Gingras, A.C. et al. (1999) Genes Dev. 13, 14221437.