Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the eIF4E translation initiation factor. 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 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).
Applications:
Suitable for use in Immunofluorescence, Flow Cytometry, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
Immunofluorescence (IF-IC): 1:400
Flow Cytometry: 1:200
Western Blot: 1:1000
Immunoprecipitation: 1:50
Immunohistochemistry (Paraffin): 1:600
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.