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PhosphoSitePlus® Resource

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REACTIVITY
Product Includes Volume Solution Color
4E-BP1 (S65) Ab Coated Microwells 96 tests
Phospho-4E-BP1 (Ser65) Rabbit Detection mAb (Biotinylated) 1 ea Green (Lyophilized)
HRP-Linked Streptavidin (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

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.


Specificity / Sensitivity

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.


Species predicted to react based on 100% sequence homology: Human

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, 762-7.

2.  Brunn, G.J. et al. (1997) Science 277, 99-101.

3.  Gingras, A.C. et al. (1998) Genes Dev 12, 502-13.

4.  Fadden, P. et al. (1997) J Biol Chem 272, 10240-7.

5.  Gingras, A.C. et al. (1999) Genes Dev 13, 1422-37.


Entrez-Gene Id 1978
Swiss-Prot Acc. Q13541


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.

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PathScan® Phospho-4E-BP1 (Ser65) Sandwich ELISA Kit