SignalSilence? MDR1/ABCB1 siRNA I

• 产品编号：CST-11879S      品牌：CST       原厂货号：11879S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNA干扰对照 > RNAi阳性对照
• 应用分类：

描述：

SignalSilence ® MDR1/ABCB1 siRNA I抑制人和猴MDR1/ABCB1基因表达。CST公司的 SignalSilence ® MDR1/ABCB1 siRNA I产品能够满足研究者通过RNA干扰方式特异性抑制MDR1/ABCB1基因表达的需求，这种方法通过将双链RNA分子释放到细胞内来实现基因表达的选择性沉默。所有CST的SignalSilence ® siRNA产品都是经过严格的内部测试，并已通过western方法证明了目标蛋白表达量的降低。

通过三苯分析监测寡核苷酸合成中的每一个碱基，以确保最佳的耦合效率。随后，该寡核苷酸经亲和固相萃取方式提纯。退火的RNA双链进一步通过质谱法进行分析，验证双链的确切组成。每个批次的谱图都会与之前批次的谱图进行比对，最大程度确保批间一致性。

CST 建议使用100 nM SignalSilence® MDR1/ABCB1 siRNA I进行转染48至72小时，之后裂解细胞。转染程序按相应转染试剂制造商提供的试验流程进行。使用过程中有任何问题，请随时联系CST。每管中含有100的转染量，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。

1. Furuya, K.N. et al. (1997) Cancer Res 57, 3708-16.
2. Litman, T. et al. (1997) Biochim Biophys Acta 1361, 169-76.
3. Chen, C.J. et al. (1986) Cell 47, 381-9.
4. Kartner, N. et al. (1983) Cancer Res 43, 4413-9.
5. Chen, G. et al. (1997) J Biol Chem 272, 5974-82.
6. Brinkmann, U. and Eichelbaum, M. (2001) Pharmacogenomics J 1, 59-64.
7. Fromm, M.F. (2004) Trends Pharmacol Sci 25, 423-9.
8. Miller, D.S. et al. (2008) Pharmacol Rev 60, 196-209.
9. Ambudkar, S.V. et al. (1999) Annu Rev Pharmacol Toxicol 39, 361-98.
10. Raviv, Y. et al. (2000) FASEB J 14, 511-5.
11. Meijer, O.C. et al. (2003) J Endocrinol 178, 13-8.
12. Karssen, A.M. et al. (2002) J Endocrinol 175, 251-60.
13. Jeannesson, E. et al. (2009) Clin Chim Acta 403, 198-202.

MDR1/ABCB1属于ATP结合盒转运蛋白超家族（1）Mdr/Tap亚家族。MDR1作为外源性化合物外排泵具有广泛的底物特异性。它的底物包括治疗剂，如放线菌素D，鬼臼乙叉甙，伊马替尼，阿霉素，以及内源性分子，如β -淀粉样蛋白，类固醇激素，脂质，磷脂，胆固醇和细胞因子（2）。研究表明， MDR1降低了肿瘤细胞中药物的累积，允许药物抗性的发展（3-5）。另一方面，在血-脑，血-脑脊髓液或血液胎盘屏障等器官中，细胞质膜MDR1的表达，限制了药物从顶或浆膜侧到这些器官中的的通透性（6,7）。MDR1基因也在正常组织如小肠，肝脏和肾脏中表达（7）以参与排泄功能。已检测到MDR1在胞内的内质网，囊泡和核膜中有表达，与细胞转运机制相关（8）。 其他已报道的MDR1的功能包括：病毒耐性，细胞因子的转运（9,10），外周和中枢神经系统的脂质平衡（11-13）。

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence® MDR1/ABCB1 siRNA I inhibits human and monkey MDR1/ABCB1 expression.

Description

SignalSilence® MDR1/ABCB1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit MDR1/ABCB1 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® MDR1/ABCB1 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

MDR1/ABCB1 belongs to the Mdr/Tap subfamily of the ATP binding casette transporter superfamily (1). MDR1 serves as an efflux pump for xenobiotic compounds with broad substrate specificity. Its substrates include therapeutic agents, such as actinomycin D, etoposide, imatinib, and doxorubicin, as well as endogenous molecules, such as β-amyloids, steroid hormones, lipids, phospholipids, cholesterol, and cytokines (2). Research studies have shown that MDR1 reduces drug accumulation in cancer cells, allowing the development of drug resistance (3-5). On the other hand, MDR1 expressed in the plasma membrane of cells in the blood-brain, blood-cerebral spinal fluid, or blood-placenta barriers restricts the permeability of drugs into these organs from the apical or serosal side (6,7). MDR1 is also expressed in normal tissues with excretory function such as small intestine, liver, and kidney (7). Intracellular MDR1 has been detected in the ER, vesicles, and nuclear envelope, and has been associated with cell trafficking machinery (8). Other reported functions of MDR1 include viral resistance, cytokine trafficking (9,10), and lipid homeostasis in the peripheral and central nervous system (11-13).

1. Furuya, K.N. et al. (1997) Cancer Res 57, 3708-16.
2. Litman, T. et al. (1997) Biochim Biophys Acta 1361, 169-76.
3. Chen, C.J. et al. (1986) Cell 47, 381-9.
4. Kartner, N. et al. (1983) Cancer Res 43, 4413-9.
5. Chen, G. et al. (1997) J Biol Chem 272, 5974-82.
6. Brinkmann, U. and Eichelbaum, M. (2001) Pharmacogenomics J 1, 59-64.
7. Fromm, M.F. (2004) Trends Pharmacol Sci 25, 423-9.
8. Miller, D.S. et al. (2008) Pharmacol Rev 60, 196-209.
9. Ambudkar, S.V. et al. (1999) Annu Rev Pharmacol Toxicol 39, 361-98.
10. Raviv, Y. et al. (2000) FASEB J 14, 511-5.
11. Meijer, O.C. et al. (2003) J Endocrinol 178, 13-8.
12. Karssen, A.M. et al. (2002) J Endocrinol 175, 251-60.
13. Jeannesson, E. et al. (2009) Clin Chim Acta 403, 198-202.

注意事项：

Storage: MDR1/ABCB1 siRNA I is supplied in RNAse-free water. Aliquot and store at -20ºC.
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