SignalSilence? Bub1b siRNA I

• 产品编号：CST-6623S      品牌：CST       原厂货号：6623S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® Bub1b siRNA能够抑制人和猴的Bub1b表达。Cell Signaling Technology (CST) 公司的SignalSilence® Bub1b siRNA 帮助研究人员使用RNA干扰技术特异性抑制Bub1b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耦合效率。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® Bub1b 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。有丝分裂检验点复合物（MCC），包括Bub1, Bub1b, Bub3, Mad2, 和Cdc20, 控制染色体分离并监控着丝粒微管相互作用（1）。在有丝分裂期，MCC复合物抑制细胞分裂后期促进复合物/ Cyclosome（APC/C）的泛素连接酶功能，从而防止未成熟折叠形成染色体的细胞进入有丝分裂后期（2）。研究证实在包括血癌，大肠癌，肺癌，乳腺癌再累的多种人类恶性肿瘤中都发现Bub1b和Bub1激酶发生了突变（3）。mosaic variegated aneuploidy 综合征和 premature chromatid separation 综合征病人也发现Bub1b基因存在双灯位基因突变（4）。在小鼠生殖细胞中敲除Bub1b基因是胚胎致死的，杂合子动物则表现出遗传不稳定性，早衰及患癌的风险上升等特点（5）。Bub3能够与Bub1和Bub1b结合，促进它们集中到着丝粒（6），在微管-着丝粒的相互作用过程中，Bub3也是不可或缺的（7）。

1. Fukagawa, T. (2008) Front Biosci 13, 2705-13.
2. Chen, R.H. (2007) J Biomed Sci 14, 475-9.
3. Dai, W. et al. (2004) Cancer Res 64, 440-5.
4. Kops, G.J. et al. (2005) Nat Rev Cancer 5, 773-85.
5. Baker, D.J. et al. (2004) Nat Genet 36, 744-9.
6. Taylor, S.S. et al. (1998) J Cell Biol 142, 1-11.
7. Logarinho, E. et al. (2008) Mol Biol Cell 19, 1798-813.

原厂资料：

omology

Species predicted to react based on 100% sequence homology: Monkey

Specificity / Sensitivity

SignalSilence® Bub1b siRNA I inhibits human and monkey Bub1b expression.

Description

SignalSilence® Bub1b siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Bub1b 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® Bub1b 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. 
 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

The Mitotic Checkpoint Complex (MCC), which contains Bub1, Bub1b, Bub3, Mad2, and Cdc20, controls chromosome segregation and monitors kinetochore-microtubule interactions (1). During mitosis, the MCC complex inhibits the ubiquitin ligase activity of the Anaphase Promoting Complex/Cyclosome (APC/C), thereby preventing cells with unaligned chromosomes from prematurely entering anaphase (2). Research studies have shown that Bub1b and Bub1 kinases are mutated in several types of human malignancies including hematopoietic, colorectal, lung, and breast cancers (3). Biallelic mutations in Bub1b have been found in mosaic variegated aneuploidy syndrome and premature chromatid separation syndrome (4). Bub1b mouse germline knockouts are embryonic lethal with heterozygous animals displaying genetic instability, early aging phenotypes, and increased cancer susceptibility (5). Bub3 binds both Bub1 and Bub1b, facilitating their recruitment to kinetochores (6), and is required for functional microtubule-kinetochore interactions (7).

1. Fukagawa, T. (2008) Front Biosci 13, 2705-13.
2. Chen, R.H. (2007) J Biomed Sci 14, 475-9.
3. Dai, W. et al. (2004) Cancer Res 64, 440-5.
4. Kops, G.J. et al. (2005) Nat Rev Cancer 5, 773-85.
5. Baker, D.J. et al. (2004) Nat Genet 36, 744-9.
6. Taylor, S.S. et al. (1998) J Cell Biol 142, 1-11.
7. Logarinho, E. et al. (2008) Mol Biol Cell 19, 1798-813.

注意事项：

Storage: Bub1b siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.