SignalSilence? LRP6 siRNA I

• 产品编号：CST-8650S      品牌：CST       原厂货号：8650S
• 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)的SignalSilence^® LRP6 siRNA I可以帮助研究者通过RNA干扰特异性地抑制 LRP6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^® LRP6 siRNA I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。LRP5和LRP6是单次跨膜蛋白，隶属于低密度脂蛋白受体（LDLR）相关蛋白家族。不同于其他LDLR家族成员，LRP5和LRP6胞外域有4个EGF和3个LDLR重复序列，胞浆域富含脯氨酸（1）。它们在经典Wnt/β-catenin信号通路中作为Wnt的共同受体（2,3）。LRP5和LRP6高度同源并且在发育过程中有多种功能（4,5）。LRP5和LRP6的活性在与Dickkopf (DKK) 家族某些蛋白结合后将被抑制（6,7）。Wnt信号通路被激活后，LRP6可以被如GSK-3和CK1之类的激酶在多个位点被磷酸化，例如1479位苏氨酸，1490位丝氨酸和1493位苏氨酸（8-10）。磷酸化的LRP6招募axin蛋白到膜上以激活β-catenin信号通路（8-10）。

1. Brown, S.D. et al. (1998) Biochem. Biophys. Res. Commun. 248, 879-888.
2. Pinson, K.I. et al. (2000) Nature 407, 535-538.
3. Tamai, K. et al. (2000) Nature 407, 530-535.
4. Kelly, O.G. et al. (2004) Development 131, 2803-2815.
5. He, X. et al. (2004) Development 131, 1663-1677.
6. Semënov, M.V. et al. (2001) Curr Biol 11, 951-61.
7. Bafico, A. et al. (2001) Nat. Cell Biol. 3, 683-668.
8. Tamai, K. et al. (2004) Mol. Cell 13, 149-156.
9. Zeng, X. et al. (2005) Nature 438, 873-877.
10. Davidson, G. et al. (2005) Nature 438, 867-872.

原厂资料：

Description

SignalSilence® LRP6 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit LRP6 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® LRP6 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.

Background

LRP5 and LRP6 are single-pass transmembrane proteins belonging to the low-density lipoprotein receptor (LDLR)-related protein family. Unlike other members of the LDLR family, LRP5 and LRP6 have four EGF and three LDLR repeats in the extracellular domain, and proline-rich motifs in the cytoplasmic domain (1). They function as co-receptors for Wnt and are required for the canonical Wnt/β-catenin signaling pathway (2,3). LRP5 and LRP6 are highly homologous and have redundant roles during development (4,5). The activity of LRP5 and LRP6 can be inhibited by the binding of some members of the Dickkopf (DKK) family of proteins (6,7). Upon stimulation with Wnt, LRP6 is phosphorylated at multiple sites including Thr1479, Ser1490, and Thr1493 by kinases such as GSK-3 and CK1 (8-10). Phosphorylated LRP6 recruits axin to the membrane and presumably activates β-catenin signaling (8-10).

1. Brown, S.D. et al. (1998) Biochem. Biophys. Res. Commun. 248, 879-888.
2. Pinson, K.I. et al. (2000) Nature 407, 535-538.
3. Tamai, K. et al. (2000) Nature 407, 530-535.
4. Kelly, O.G. et al. (2004) Development 131, 2803-2815.
5. He, X. et al. (2004) Development 131, 1663-1677.
6. Semënov, M.V. et al. (2001) Curr Biol 11, 951-61.
7. Bafico, A. et al. (2001) Nat. Cell Biol. 3, 683-668.
8. Tamai, K. et al. (2004) Mol. Cell 13, 149-156.
9. Zeng, X. et al. (2005) Nature 438, 873-877.
10. Davidson, G. et al. (2005) Nature 438, 867-872.

注意事项：

Storage: LRP6 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.