PathScan® Phospho-4E-BP1 (Ser65) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of 4E-BP1 when phosphorylated at Ser65. A total 4E-BP1 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, 4E-BP1 protein is captured by the coated antibody. Following extensive washing, a biotinylated Phospho-4E-BP1 (Ser65) Rabbit mAb Detection Antibody is added to detect the captured phospho-4E-BP1 protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of 4E-BP1 phosphorylated at Ser65.
Antibodies in kit are custom formulations specific to kit.
PathScan® Phospho-4E-BP1 (Ser65) Sandwich ELISA Kit detects endogenous levels of 4E-BP1 protein when phosphorylated at Ser65, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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