SignalSilence? Sec24B siRNA II

• 产品编号：CST-12333S      品牌：CST       原厂货号：12333S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence^® Sec24B siRNA II抑制人和猴中Sec24B的表达。来自Cell Signaling Technology (CST)的SignalSilence^® Sec24B siRNA II可以帮助研究者通过RNA干扰特异性地抑制Sec24B 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^®Sec24B 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。外壳蛋白复合物Ⅱ（COPⅡ）由5种胞浆蛋白组成：Sec23/24 复合物, Sec13/31 复合物, 和 Sar1. COPII外壳位于内质网/高尔基体的接口处，设计到新和成的蛋白从内质网向高尔基体的转运（1）。COPⅡ的形成起始于激活的G蛋白，Sar1与Sec23/24复合物结合，随后形成一个前萌芽复合物直接与靶分子结合（1-3）。前萌芽复核无误随后募集Sec13/31形成成熟的外壳蛋白复合物Ⅱ（4,5）。外壳蛋白复合物Ⅱ的Sec24亚基被认为在cargo选择过程中发挥了关键性左右（2,6）。它和cargo蛋白在内质网中直接结合并通过Sec23将他们带到COPII 囊泡。人细胞中有4种Sec24亚型：Sec24A, Sec24B, Sec24C, 和Sec24D (7).小鼠中，Sec24B的突变与多种发育缺陷相关（8,9）。

1. Aridor, M. et al. (1998) J Cell Biol 141, 61-70.
2. Miller, E.A. et al. (2003) Cell 114, 497-509.
3. Mossessova, E. et al. (2003) Cell 114, 483-95.
4. Barlowe, C. et al. (1994) Cell 77, 895-907.
5. Bi, X. et al. (2007) Dev Cell 13, 635-45.
6. Miller, E. et al. (2002) EMBO J 21, 6105-13.
7. Tang, B.L. et al. (1999) Biochem Biophys Res Commun 258, 679-84.
8. Merte, J. et al. (2010) Nat Cell Biol 12, 41-6; sup pp 1-8.
9. Wansleeben, C. et al. (2010) Development 137, 1067-73.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® Sec24B siRNA II inhibits human and monkey Sec24B expression.

Description

SignalSilence® Sec24B siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Sec24B 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® Sec24B 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

Coat Protein Complex II (COPII) is composed of five cytosolic proteins: Sec23/24 complex, Sec13/31 complex, and Sar1. COPII coat is located at the ER/Golgi interface and is involved in transport of newly synthesized proteins from the ER to the Golgi apparatus (1). COPII formation is initiated through the binding of the activated G protein, Sar1, to the Sec23/24 complex, thereby forming a prebudding complex that directly binds target molecules (1-3). The prebudding complex further recruits Sec13/31 to form mature COPII coat (4,5). The Sec24 subunit of COPII coat is thought to play a critical role in cargo selection (2,6). It binds directly to cargo proteins at the ER and brings them to COPII vesicles through interaction with Sec23. There are four Sec24 isoforms in human cells: Sec24A, Sec24B, Sec24C, and Sec24D (7). In mice, mutations in Sec24B have been linked to developmental defects (8,9).

1. Aridor, M. et al. (1998) J Cell Biol 141, 61-70.
2. Miller, E.A. et al. (2003) Cell 114, 497-509.
3. Mossessova, E. et al. (2003) Cell 114, 483-95.
4. Barlowe, C. et al. (1994) Cell 77, 895-907.
5. Bi, X. et al. (2007) Dev Cell 13, 635-45.
6. Miller, E. et al. (2002) EMBO J 21, 6105-13.
7. Tang, B.L. et al. (1999) Biochem Biophys Res Commun 258, 679-84.
8. Merte, J. et al. (2010) Nat Cell Biol 12, 41-6; sup pp 1-8.
9. Wansleeben, C. et al. (2010) Development 137, 1067-73.

注意事项：

Storage: Sec24B siRNA II is supplied in RNAse-free water.
Aliquot and store at -20ºC