Cell Signaling Technology

Product Pathways - Translational Control

SignalSilence® hnRNP E1 siRNA II #7987

Applications Reactivity
Transfection H

Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® hnRNP E1 siRNA I #7798 (+) or SignalSilence® hnRNP E1 siRNA II (+), using hnRNP E1 Antibody #8534 (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The hnRNP E1 Antibody confirms silencing of hnRNP E1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Description

SignalSilence® hnRNP E1 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit hnRNP E1 expression using 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 products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.CST recommends transfection with 100 nM SignalSilence® hnRNP E1 siRNA I 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.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® hnRNP E1 siRNA II 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.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

hnRNP E1 is a member of the hnRNP family of proteins that are involved in pre-mRNA processing and mRNA export, localization, stability, and translation (1-6). hnRNP E1 exerts a wide range of biological functions, such as transcriptional activation of mouse MOR gene expression (7), attenuation of alternative splicing of GHR pseudoexon expression (8), stabilization of collagen I and II (9), beta-globin (10), and androgen receptor (11) mRNAs, and regulation of translation of various genes including Dab2, ILEI, and Bag-1 (12,13). hnRNP E1 is ubiquitously expressed. Phosphorylation of hnRNP E1 affects its RNA binding affinity (13,14).

  1. Cáceres, J.F. et al. (1994) Science 265, 1706-9.
  2. Gallouzi, I.E. and Steitz, J.A. (2001) Science 294, 1895-901.
  3. Carson, J.H. et al. (2001) Results Probl Cell Differ 34, 69-81.
  4. Xu, K. et al. (2001) J Immunol 166, 6358-66.
  5. Habelhah, H. et al. (2001) Nat Cell Biol 3, 325-30.
  6. Dreyfuss, G. et al. (2002) Nat Rev Mol Cell Biol 3, 195-205.
  7. Kim, S.S. et al. (2005) Mol Pharmacol 68, 729-36.
  8. Akker, S.A. et al. (2007) Mol Endocrinol 21, 2529-40.
  9. Thiele, B.J. et al. (2004) Circ Res 95, 1058-66.
  10. Yu, J. and Russell, J.E. (2001) Mol Cell Biol 21, 5879-88.
  11. Yeap, B.B. et al. (2002) J Biol Chem 277, 27183-92.
  12. Pickering, B.M. et al. (2003) Nucleic Acids Res 31, 639-46.
  13. Chaudhury, A. et al. (2010) Nat Cell Biol 12, 286-93.
  14. Leffers, H. et al. (1995) Eur J Biochem 230, 447-53.

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.

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

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