Cell Signaling Technology

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SignalSilence® ILK1 siRNA (Human Specific) #6202

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with 50 nM non-targeted siRNA (-) or 100 nM ILK1 siRNA (+), using ILK1 Antibody #3862 and Cofilin Antibody #3312. The ILK1 Antibody confirms silencing of ILK1 expression, and the Cofilin Antibody controls for protein loading and ILK1 siRNA specificity.

IF-IC

IF-IC

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

CST recommends transfection with 100 nM human-specific ILK1 siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.

Background

Integrin-linked kinases (ILKs) couple integrins and growth factors to downstream pathways involved in cell survival, cell cycle control, cell-cell adhesion and cell motility (1). ILK functions as a scaffold bridging the extracellular matrix (ECM) and growth factor receptors to the actin cytoskeleton through interactions with integrin, PINCH (which links ILK to the RTKs via Nck2), CH-ILKBP and affixin (1). ILK phosphorylates Akt at Ser473, GSK-3 on Ser9, myosin light chain 2 (MLC2) on Ser18/Thr19, as well as affixin (2-5). These phosphorylation events are key regulatory steps in modulating the activities of the targets. ILK activity is stimulated by PI3 kinase and negatively regulated by the tumor suppressor PTEN and a PP2C protein phosphatase, ILKAP (1,3,6). It has been suggested that the conserved Ser343 residue in the activation loop plays a key role in the activation of ILK1 (2).

Small Interfering RNA (siRNA) has been used to specifically silence ILK expression in HEK-293 cells (7).

  1. Wu, C. and Dedhar, S. (2000) J. Biol. Chem. 155, 505-510.
  2. Persad, S. et al. (2001) J. Biol. Chem. 276, 27462-27469.
  3. Persad, S. et al. (2000) J. Cell Biol. 153, 1161-1173.
  4. Deng, J.T. et al. (2001) J. Biol. Chem. 276, 16365-16373.
  5. Yamaji, S. et al. (2001) J. Cell Biol. 153, 1251-1264.
  6. Morimoto, A.M. et al. (2000) Oncogene 19, 200-209.
  7. Troussard, A. A. et al. (2003) J. Biol. Chem. 278(25) , 22374-22378.

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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