Cell Signaling Technology

Product Pathways - Translational Control

SignalSilence® eIF4E siRNA #6311

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untransfected or transfected with either nonspecific control siRNA or eIF4E siRNA. eIF4E was detected using eIF4E Antibody #9742, and eIF4B was detected using eIF4B Antibody #3592. The eIF4E Antibody confirms silencing of eIF4E expression, and the eIF4B Antibody is used to control for loading and siRNA specificity.

Fluorescent Detection

Fluorescent Detection

Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.

Directions for Use

Background

Eukaryotic initiation factor 4E (eIF4E) binds to the mRNA cap structure to mediate the initiation of translation (1,2). eIF4E interacts with eIF4G, a scaffold protein that promotes assembly of eIF4E and eIF4A into the eIF4F complex (2). eIF4B is thought to assist the eIF4F complex in translation initiation. Upon activation by mitogenic and/or stress stimuli mediated by Erk and p38 MAPK, Mnk1 phosphorylates eIF4E at Ser209 in vivo (3,4). Two Erk and p38 MAPK phosphorylation sites in mouse Mnk1 (Thr197 and Thr202) are essential for Mnk1 kinase activity (3). The carboxy-terminal region of eIF4G also contains serum-stimulated phosphorylation sites, including Ser1108, Ser1148 and Ser1192 (5). Phosphorylation at these sites is blocked by the PI3 kinase inhibitor LY294002 and by the FRAP/mTOR inhibitor rapamycin.

  1. Sonenberg, N. et al. (1978) Proc. Natl. Acad. Sci. USA 75, 4843-4847.
  2. Gingras, A.C. et al. (1999) Annu. Rev. Biochem. 68, 913-963.
  3. Waskiewicz, A. et al. (1999) Mol. Cell. Biol. 19, 1871-1880.
  4. Pyronnet, S. et al. (1999) EMBO J. 18, 270-279.
  5. Raught, B. et al. (2000) EMBO J. 19, 434-444.
  6. Yamagiwa , Y. et al. (2003) Hepatology 38, 158-166.

Application References

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Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

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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