Cell Signaling Technology

Product Pathways - Translational Control

4E-BP1 Blocking Peptide #1053

Description

This peptide is used to block 4E-BP1 (53H11) Rabbit mAb #9644 reactivity in immunohistochemistry protocols, and #9644 and 9452 reactivity in western blot protocols.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks 4E-BP1 (53H11) Rabbit mAb #9644 signal in immunohistochemistry.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using 4E-BP1 (53H11) Rabbit mAb #9644 in the presence of control peptide (left) or 4E-BP1 Blocking Peptide (right).

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 μl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the product data sheet.

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, 762-767.
  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.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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