SignalSilence? PI3 Kinase p110β siRNA I

• 产品编号：CST-13364S      品牌：CST       原厂货号：13364S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST) 公司的PI3 Kinase p110β siRNA I帮助研究人员使用RNA干扰技术特异性抑制PI3 Kinase p110β表达，该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试，western分析显示能够能够降低目标蛋白的表达。寡核苷酸合成经过了三苯甲基的逐碱基分析以确保偶联效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。CST建议转染100 nM SignalSilence® PI3 Kinase p110β siRNA I，48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题，可以随时咨询CST。 每管试剂能进行100次转染，该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。PI3K激酶通过磷酸化磷脂酰肌醇(PI), 4-磷酸磷脂酰肌醇 (PIP)和4，5-二磷酸磷脂酰肌醇 (PIP2)催化3,4,5-三磷酸磷脂酰肌醇的产生。生长因子和激素会引发该磷酸化进程，随后协助细胞生长，细胞周期进入，细胞迁移和细胞存活（1）。PTEN会逆转该过程，研究表明PI3K信号通路在PTEN功能缺失的人类癌症中是持续性激活的（2）。PI3Ks由一个催化亚基（p110）和一个调控亚基构成。已经分离得到了多个催化亚基(p110α, p110β, p110γ, 和p110δ)，与p110α, p110β, 和 p110δ结合的调控亚基是p85α 和 p85β (3).相对的，与p101调控亚基结合的p110γ与p85无关。此外，异源三聚体G蛋白的βγ亚基可以激活p110γ(4)。此外，异源三聚体G蛋白的βγ亚基可以激活p110γ(4)。p110β激酶亚基在多种组织中表达并在胚胎早期发育中发挥了重要作用（5）。p110β刺激细胞增殖和侵润细胞生长；在多种肿瘤包括恶性胶质瘤中表达上升（6-8）。

1. Cantley, L.C. (2002) Science 296, 1655-7.
2. Simpson, L. and Parsons, R. (2001) Exp Cell Res 264, 29-41.
3. Neri, L.M. et al. (2002) Biochim Biophys Acta 1584, 73-80.
4. Stoyanov, B. et al. (1995) Science 269, 690-3.
5. Okkenhaug, K. and Vanhaesebroeck, B. (2003) Nat Rev Immunol 3, 317-30.
6. Czauderna, F. et al. (2003) Nucleic Acids Res 31, 670-82.
7. Bénistant, C. et al. (2000) Oncogene 19, 5083-90.
8. Knobbe, C.B. and Reifenberger, G. (2003) Brain Pathol 13, 507-18.

原厂资料：

Description

SignalSilence^® PI3 Kinase p110β siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PI3 Kinase p110β 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 CST™ SignalSilence^® siRNA products 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 transfecting with 100 nM PI3 Kinase p110β siRNA I 48 to 72 hours before lysing cells. 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

Phosphoinositide 3-kinase (PI3K) catalyzes the production of phosphatidylinositol-3,4,5-triphosphate by phosphorylating phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol-4,5-bisphosphate (PIP₂). Growth factors and hormones trigger this phosphorylation event, which in turn coordinates cell growth, cell cycle entry, cell migration, and cell survival (1). PTEN reverses this process, and research studies have shown that the PI3K signaling pathway is constitutively activated in human cancers that have loss of function of PTEN (2). PI3Ks are composed of a catalytic subunit (p110) and a regulatory subunit. Various isoforms of the catalytic subunit (p110α, p110β, p110γ, and p110δ) have been isolated, and the regulatory subunits that associate with p110α, p110β, and p110δ are p85α and p85β (3). In contrast, p110γ associates with a p101 regulatory subunit that is unrelated to p85. Furthermore, p110γ is activated by βγ subunits of heterotrimeric G proteins (4).The p110β kinase subunit has a broad tissue distribution and plays an essential role in early embryonic development (5). p110β stimulates cell proliferation and invasive cell growth; expression is increased in a number of tumors, including glioblastomas (6-8).

1. Cantley, L.C. (2002) Science 296, 1655-7.
2. Simpson, L. and Parsons, R. (2001) Exp Cell Res 264, 29-41.
3. Neri, L.M. et al. (2002) Biochim Biophys Acta 1584, 73-80.
4. Stoyanov, B. et al. (1995) Science 269, 690-3.
5. Okkenhaug, K. and Vanhaesebroeck, B. (2003) Nat Rev Immunol 3, 317-30.
6. Czauderna, F. et al. (2003) Nucleic Acids Res 31, 670-82.
7. Bénistant, C. et al. (2000) Oncogene 19, 5083-90.
8. Knobbe, C.B. and Reifenberger, G. (2003) Brain Pathol 13, 507-18.

注意事项：

Storage: PI3 Kinase p110β siRNA is supplied in RNAse-free
water. Aliquot and store at -20ºC.