New England Biolabs Canada
 

Product Pathways - Protein Translation

Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855

Item# Description List Price Web Price Qty
2855L Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb - 300 µl $1,057.00
$951.30
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2855S Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb - 100 µl $446.00
$401.40
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2855T Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb - 20 µl $180.00
$162.00
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Monkey, D. melanogaster Endogenous 15 to 20 Rabbit IgG
IHC-P
IF-IC
F

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry

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. Non-specific staining has been observed in mitotic cells by immunofluorescence.

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 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)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.


IHC-P (paraffin)

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)

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).

IF-IC

IF-IC

Confocal immunofluorescent analysis of 293 cells, expressing either non-targeting shRNA (top) or shRNA targeting 4E-BP1/2 (bottom), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). To confirm phospho-specificity, cells were treated with an inhibitor cocktail consisting of LY294002 #9901, U0126 #9903, and Rapamycin #9904 (50 μM; 10 μm; 10 nM; 2 hr) (left), stimulated with insulin (100 nM, 30 min; middle), or processed with λ-phosphatase following insulin stimulation (right). Red = Propidium Iodide (PI)/RNase Staining Solution (#4087).


Flow Cytometry

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).

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).

  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.

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.
  • Bu, Y. et al. (2018) Nat Cell Biol 20, 104-115. 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!


 

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