Cell Signaling Technology

Product Pathways - Translational Control

4E-BP2 Antibody #2845

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P IF-IC F H M R Mk Pg B Endogenous 15 to 20 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig  B=Bovine
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

4E-BP2 Antibody detects endogenous levels of total 4E-BP2, independent of phosphorylation. This antibody does not cross-react significantly with 4E-BP1.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues at the carboxy-terminus of human 4E-BP2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from A673 cells, untreated or nocodazole-treated (100 ng/ml,16hrs), using 4E-BP2 Antibody (upper) or 4E-BP1 Antibody #9452 (lower). Extracts were treated with lambda phosphatase NEB#P0753 (10,000 U/ml for 1 hour) to dephosphorylate both proteins.

Western Blotting

Western Blotting

Western blot analysis of bacterially expressed GST-4E-BP1 and of extracts from NIH/3T3 cells, using 4E-BP2 Antibody and 4E-BP1 Antibody #9452.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic and nuclear localization, using 4E-BP2 Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using 4E-BP2 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human follicular carcinoma (thyroid), using 4E-BP2 Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, using 4E-BP2 Antibody (blue) compared to a nonspecifc negative control antibody (red).


IF-IC

IF-IC

Immunofluorescent detection of 4E-BP2 (green) in A673 cells, using 4E-BP2 Antibody (left). Brightfield image is shown (right).

Background

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).

4E-BP2 and 4E-BP3 share high sequence homology with 4E-BP1, including conservation of the major FRAP/mTOR-dependent phosphorylation sites. Preliminary data suggests that phosphorylation of 4E-BP2 is regulated in a similar manner to that of 4E-BP1, although phosphorylation of this protein has not been as extensively studied (6).

  1. Pause, A. et al. (1994) Nature 371, 762-767.
  2. Brunn, G.J. et al. (1997) Science 277, 99-101.
  3. Gingras, A.C. et al. (1998) Genes Dev. 12, 502-513.
  4. Fadden, P. et al. (1997) J. Biol. Chem. 272, 10240-10247.
  5. Gingras, A.C. et al. (1999) Genes Dev. 13, 1422-1437.
  6. Lin, T.A. and Lawrence, Jr, J.C. (1996) J. Biol. Chem. 271, 30199-30204.

Application References

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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