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51367
4E-BP1 Control Cell Extracts
Experimental Controls
Cell Extract Kit

4E-BP1 Control Cell Extracts #51367

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  1. WB
Western Blotting Image 1: 4E-BP1 Control Cell Extracts

Western blot analysis of 4E-BP1 Control Cell Extracts from MCF7 cells, amino acid starved (1 hr), then either untreated (-) or treated with insulin (100 nM, 30 min; +), using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855, Phospho-4E-BP1 (Ser65) (174A9) Rabbit mAb #9456, Phospho-4E-BP1 (Thr70) Antibody #9455, Phospho-4E-BP1 (Ser65) (D9G1Q) Rabbit mAb #13443, and Phospho-4E-BP1 (Thr37/46) Antibody #9459.

Product Usage Information

Boil for 3 minutes prior to use. Load 10 μl of phosphorylated and nonphosphorylated Smad2/3 Control Cell Extracts per lane.

Storage

Store at -20°C. Supplied in SDS Sample Buffer: 62.5 mM Tris- HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM DTT, 0.01% w/v bromophenol blue or phenol red.

Product Description

Nonphosphorylated 4E-BP1 Control Cell Extracts: Total cell extracts from MCF7 cells, amino acids starved for 1 hour to serve as a negative control. Supplied in SDS Sample Buffer.

Phosphorylated 4E-BP1 Control Cell Extracts: Total cell extracts from MCF7 cells, amino acids starved for 1 hour followed by adding back amino acids for 1 hour and treating with 100 nM insulin for 30 min to serve as a positive control. Supplied in SDS Sample Buffer.

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.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
To Purchase # 51367S
Product # Size Price
51367S
100 µl  (10 western blots) N/A

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