Product Pathways - Adhesion
SignalSilence® ILK1 siRNA Kit (Human Specific) #6200
Kit Includes
- SignalSilence® ILK1 siRNA (Human Specific) #6202
- pre-validated single sequence siRNA duplex guaranteed to specifically inhibit ILK1 expression in specified cell lines.
- SignalSilence® Control siRNA (Fluorescein Conjugate) #6201
- a non-targeted negative control duplex designed to monitor transfection efficiency using microscopy and to monitor siRNA specificity.
- Targeted ILK1 Antibody #3862
- to confirm silencing of ILK1 expression.
- Non-targeted Cofilin Antibody #3312
- to control for protein loading and ILK1 siRNA specificity.
- *Transfection Reagent
- designed specifically for highly efficient siRNA delivery in mammalian cells. Transfection reagent is supplied by Mirus.
*Store at 4°C (packaged separately)
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.
Fluorescent Detection
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
Description
SignalSilence® ILK1 siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit ILK1 expression. The kit utilizes RNA interference, a method in which gene expression can be selectivelysilenced 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® ILK1 siRNA Kit includes ILK1 siRNA, a target-specific ILK1 Antibody to confirm the silencing of ILK1 expression and a non-target antibody (Cofilin) to control for loading and monitor specificity of ILK1 siRNA. Additionally, a fluorescein-labeled non-targeted siRNA control allows the user to monitor transfection efficiency and siRNA specificity. The kit also contains a transfection reagent designed solely for efficient siRNA delivery in mammalian cells.
Directions for Use
CST recommends transfection with 100 nM human-specific ILK1 siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.
Tested Cell Lines
HeLa
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).
- Wu, C. and Dedhar, S. (2000) J. Biol. Chem. 155, 505-510.
- Persad, S. et al. (2001) J. Biol. Chem. 276, 27462-27469.
- Persad, S. et al. (2000) J. Cell Biol. 153, 1161-1173.
- Deng, J.T. et al. (2001) J. Biol. Chem. 276, 16365-16373.
- Yamaji, S. et al. (2001) J. Cell Biol. 153, 1251-1264.
- Morimoto, A.M. et al. (2000) Oncogene 19, 200-209.
- Troussard, A. A. et al. (2003) J. Biol. Chem. 278(25) , 22374-22378.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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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.
