Cell Signaling Technology

Product Pathways - Translational Control

SignalSilence® eIF4E siRNA Kit #6555

Products Included No. Quantity Isotype Assay
SignalSilence® eIF4E siRNA I 6311 10 µM in 150 µl 50 Transfections
SignalSilence® eIF4E siRNA II 6554 10 µM in 150 µl 50 Transfections
eIF4E (C46H6) Rabbit mAb 2067 40 µl Rabbit IgG 4 Western mini blots

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® eIF4E siRNA I (+) or SignalSilence® eIF4E siRNA II (+), using eIF4E (C46H6) Rabbit mAb and α-Tubulin (11H10) Rabbit mAb #2125. The eIF4E (C46H6) Rabbit mAb confirms silencing of eIF4E expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of eIF4E siRNA.

Description

SignalSilence® eIF4E siRNA Kit from Cell Signaling Technology (CST) allows the researcher to specifically inhibit eIF4E expression. The kit utilizes RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA kits are rigorously tested in-house and have been shown to reduce protein expression. SignalSilence® eIF4E siRNA Kit includes eIF4E siRNA I and II and a target-specific eIF4E Antibody to confirm the silencing of eIF4E expression by western analysis.

Directions for Use

CST recommends transfection with 100 nM eIF4E siRNA 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on 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.

Application References

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Protocols

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 intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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