SignalSilence? LIMK1 siRNA II

• 产品编号：CST-7156S      品牌：CST       原厂货号：7156S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

LIMK1 siRNA I抑制人和猴中LIMK1的表达。来自Cell Signaling Technology (CST)的SignalSilence^® LIMK1 siRNA II可以帮助研究者通过RNA干扰特异性地抑制Stat6的表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。通过三苯甲基分析每个碱基以监测寡核苷酸的合成，确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较，来保证不同批次之间的最大一致性。CST推荐使用100 nM SignalSilence^® LIMK1 siRNA II 进行转染，48到72小时后对细胞进行裂解。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。每小瓶可供100次转染，每次转染量相当于在转染24孔板时，每孔总体积为300μl培养基中siRNA的终浓度为100nM。LIM激酶（LIMK1和LIMK2）是丝氨酸/苏氨酸激酶，有两个位于氨基调控末端的锌指结构域，称为LIM结构域（1）。LIM激酶涉及到肌动蛋白丝细胞骨架调节下游Rho0家族GTPases，PAKs和ROCK（2,3）。PAK1和ROCK磷酸化LIMK1或LIMK2的激活茎环保守的508位或505位苏氨酸，提升LIMK活性（3-5）。激活的LIM激酶通过磷酸化cofilin氨基末端第3位丝氨酸残基而抑制cofilin的肌动蛋白去多聚化活性（6,7）。

1. Okano, I. et al. (1995) J. Biol. Chem. 270, 31321-31330.
2. Maekawa, M. et al. (1999) Science 285, 895-898.
3. Edwards, D. C. et al. (1999) Nat. Cell Biol. 1, 253-259.
4. Ohashi, K. et al. (2000) J. Biol. Chem. 275, 3577-3582.
5. Sumi, T. et al. (2001) J. Biol. Chem. 276, 670-676.
6. Arber, S. et al. (1998) Nature 393, 805-809.
7. Yang, N. et al. (1998) Nature 393, 809-812.

原厂资料：

Specificity / Sensitivity

LIMK1 siRNA II inhibits human and monkey LIMK1 expression.

Description

SignalSilence® LIMK1 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit LIMK1 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.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® LIMK1 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

LIM kinases (LIMK1 and LIMK2) are serine/threonine kinases that have two zinc finger motifs, known as LIM motifs, in their amino-terminal regulatory domains (1). LIM kinases are involved in actin cytoskeletal regulation downstream of Rho-family GTPases, PAKs, and ROCK (2,3). PAK1 and ROCK phosphorylate LIMK1 or LIMK2 at the conserved Thr508 or Thr505 residues in the activation loop, increasing LIMK activity (3-5). Activated LIM kinases inhibit the actin depolymerization activity of cofilin by phosphorylation at the amino-terminal Ser3 residue of cofilin (6,7).

1. Okano, I. et al. (1995) J. Biol. Chem. 270, 31321-31330.
2. Maekawa, M. et al. (1999) Science 285, 895-898.
3. Edwards, D. C. et al. (1999) Nat. Cell Biol. 1, 253-259.
4. Ohashi, K. et al. (2000) J. Biol. Chem. 275, 3577-3582.
5. Sumi, T. et al. (2001) J. Biol. Chem. 276, 670-676.
6. Arber, S. et al. (1998) Nature 393, 805-809.
7. Yang, N. et al. (1998) Nature 393, 809-812.

注意事项：

Storage: LIMK1 siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC.