Product Pathways - Translational Control
Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855
PhosphoSitePlus® protein, site, and accession data: 4E-BP1
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IHC-P IF-IC F | H M R Mk Dm | Endogenous | 15 to 20 | Rabbit IgG |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Dm=D. melanogaster
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb detects endogenous levels of 4E-BP1 only when phosphorylated at Thr37 and/or Thr46. This antibody may cross-react with 4E-BP2 and 4E-BP3 when phosphorylated at equivalent sites.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr37 and Thr46 of mouse 4E-BP1.
Western Blotting
Western blot analysis of extracts from 293T cells using 4E-BP1 Antibody #9452 (upper) and Phospho-4E-BP1 (Thr37/46) Antibody #2855 (lower). 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.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells untreated (left) or LY294002-treated (right)).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb in the presence of control peptide (left) or Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (right).
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue), using Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent analysis of HeLa cells treated with LY294002 (left) or 20% serum (right) and labeled with Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Background
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.
Application References
- Engelman, J.A. et al. (2008) Nat. Med. 14, 1351-1356. Applications: IHC-P (paraffin)
- Cao, R. et al. (2008) Mol. Cell. Neurosci. 38, 312-324. Applications: IHC-P (paraffin)
- Gómez-Abad, C. et al. (2011) Blood 118, 5517-27. Applications: Western Blotting
- Roczniak-Ferguson, A. et al. (2012) Sci Signal 5, ra42. Applications: Western Blotting
- Thomson, D.M. et al. (2010) J Appl Physiol 108, 1775-85. Applications: Western Blotting
- Tagoug, I. et al. (2011) PLoS One 6, e22641. Applications: Western Blotting
- Wang, Z. et al. (2011) Autophagy 7, 401-11. Applications: Western Blotting
- Murai, A. et al. (2012) BMC Vet Res 8, 128. Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9644 4E-BP1 (53H11) Rabbit mAb
- 9459 Phospho-4E-BP1 (Thr37/46) Antibody
- 9456 Phospho-4E-BP1 (Ser65) (174A9) Rabbit mAb
- 9455 Phospho-4E-BP1 (Thr70) Antibody
- 8112 SignalStain® Antibody Diluent
- 9451 Phospho-4E-BP1 (Ser65) Antibody
- 9452 4E-BP1 Antibody
- 4923 Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb
- 2845 4E-BP2 Antibody
- 9955 4E-BP Antibody Sampler Kit
- 9918 Translational Control Antibody Sampler Kit
- 1052 Phospho-4E-BP1 (Thr37/46) Blocking Peptide
- 9741 Phospho-eIF4E (Ser209) Antibody
- 9742 eIF4E Antibody
- 9862 mTOR Substrates Antibody Sampler Kit
- 7216 PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Kit
- 8101 SignalSlide® Phospho-Akt (Ser473) IHC Controls
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
- 5425 Normal Goat Serum
Rabbit monoclonal antibody is produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and U.S.S.N. 11/476,277) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.
