SignalSilence? ILK1 siRNA II

• 产品编号：CST-6528S      品牌：CST       原厂货号：6528S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

来自Cell Signaling Technology (CST)公司的SignalSilence® ILK1 siRNA II让研究者用RNA 干扰技术选择性抑制表达，这是一种通过发送双链RNA分子到细胞特异抑制ILK1基因表达。所有SignalSilence®siRNA产物来自CST，经过严格的内部测试，并由western analysis已经显示减少蛋白表达。寡核苷酸合成通过三苯甲基单个碱基监测，保证合适的配对效率。寡核苷酸随后由亲和固相抽提法纯化。退火的RNA双螺旋由质谱进一步分析证明双螺旋的准确组成。每一批由质谱比对前一批，以保证最大的批与批之间的一致性。CST 推荐在细胞裂解前48到72小时将100nM ILK1 siRNA II转染。对应转染步骤，按转染试剂生产商提供的步骤进行，如在使用上有任何问题请随时联系CST。整合素连接酶(ILKs)配对整合素和生长因子到下游通路涉及细胞存活，周期控制，细胞与细胞粘附和细胞移动（1）。ILK作为一个支架，通过与PINCH（此蛋白通过Nck2连接ILK到RTKs），CH-ILKBP 和affixin相互作用，桥连细胞外基质(ECM)和生长因子受体到肌动蛋白骨架（1）。ILK磷酸化Akt的473位丝氨酸，磷酸化GSK-3 的9位丝氨酸，磷酸化肌球蛋白轻链2（MLC2）的18位丝氨酸/19位苏氨酸，还有affixin（2-5）。这些磷酸化时调控靶标活性的关键调控步骤。ILK活性被PI3激酶刺激，被肿瘤抑制因子PTEN和PP2C蛋白磷酸酶ILKAP负向调控（1,3,6）。已经暗示，保守的343位丝氨酸残基在激活环中对ILK1的激活起关键作用（2）。

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.

原厂资料：

Description

SignalSilence® ILK1 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ILK1 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 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.

Directions for Use

CST recommends transfection with 100 nM ILK1 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.

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).

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.

注意事项：

Storage: ILK1 siRNA II is supplied in RNAse-free water. Aliquot
and store at -20°C.