SignalSilence? Mcl-1 siRNA I

• 产品编号：CST-6315S      品牌：CST       原厂货号：6315S
• 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^® Mcl-1 siRNA I可以帮助研究者通过RNA干扰特异性地抑制Mcl-1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^® Mcl-1 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。Mcl-1 是Bcl-2家族的抗凋亡成员，首次在佛波酯诱导单核细胞/巨噬细胞通路分化时，从ML-1人髓系白血病细胞系中被分离(1) 。类似于其他 Bcl-2家族成员，Mcl-1定位于线粒体 (2)，通过与促凋亡Bcl-2 家族成员互作或拮抗（3），抑制细胞毒性刺激诱导的细胞凋亡（4）。与其他家族成员不同，Mcl-1可以在转录和转录后修饰水平进行调控。首先，Mcl-1 有一个扩展的氨基末端PEST区域，负责其相对较短的半衰期 (1，2)。第二，与其他家族成员不同，Mcl-1通过PI3K/Akt依赖的通路迅速被转录，从而在髓细胞分化和细胞因子刺激时提高表达量 (1，5-7)。Mcl-1在用佛波酯、微管损伤试剂、氧化应激和细胞因子处理时，会被磷酸化(8-11)。在PEST区域内保守的MAP kinase/ERK位点，Thr163发生磷酸化会减缓 Mcl-1 蛋白质的转化 (10)，但可能会刺激GSK-3介导Ser159磷酸化，导致Mcl-1的不稳定 (11)。小鼠缺乏Mcl-1会导致围着床期致死(12)。此外，条件性终止相应的 mcl-1基因了Mcl-1在早期淋巴组织的发育和成熟淋巴细胞的维持过程中发挥了重要作用(13)。

1. Kozopas, K.M. et al. (1993) Proc Natl Acad Sci USA 90, 3516-20.
2. Yang, T. et al. (1995) J Cell Biol 128, 1173-84.
3. Sato, T. et al. (1994) Proc Natl Acad Sci USA 91, 9238-42.
4. Zhou, P. et al. (1997) Blood 89, 630-43.
5. Wang, J.M. et al. (1999) Mol Cell Biol 19, 6195-206.
6. Jourdan, M. et al. (2003) Oncogene 22, 2950-9.
7. Chao, J.R. et al. (1998) Mol Cell Biol 18, 4883-98.
8. Domina, A.M. et al. (2000) J Biol Chem 275, 21688-94.
9. Inoshita, S. et al. (2002) J Biol Chem 277, 43730-4.
10. Domina, A.M. et al. (2004) Oncogene 23, 5301-15.
11. Maurer, U. et al. (2006) Mol Cell 21, 749-60.
12. Rinkenberger, J.L. et al. (2000) Genes Dev 14, 23-7.
13. Opferman, J.T. et al. (2003) Nature 426, 671-6.

原厂资料：

Description

SignalSilence® Mcl-1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Mcl-1 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 Mcl-1 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

Mcl-1 is an anti-apoptotic member of the Bcl-2 family originally isolated from the ML-1 human myeloid leukemia cell line during phorbol ester-induced differentiation along the monocyte/macrophage pathway (1). Similar to other Bcl-2 family members, Mcl-1 localizes to the mitochondria (2), interacts with and antagonizes pro-apoptotic Bcl-2 family members (3), and inhibits apoptosis induced by a number of cytotoxic stimuli (4). Mcl-1 differs from its other family members in its regulation at both the transcriptional and post-translational level. First, Mcl-1 has an extended amino-terminal PEST region, which is responsible for its relatively short half-life (1,2). Second, unlike other family members, Mcl-1 is rapidly transcribed via a PI3K/Akt dependent pathway, resulting in its increased expression during myeloid differentiation and cytokine stimulation (1,5-7). Mcl-1 is phosphorylated in response to treatment with phorbol ester, microtubule-damaging agents, oxidative stress, and cytokine withdrawal (8-11). Phosphorylation at Thr163, the conserved MAP kinase/ERK site located within the PEST region, slows Mcl-1 protein turnover (10) but may prime the GSK-3 mediated phosphorylation at Ser159 that leads to Mcl-1 destabilization (11). Mcl-1 deficiency in mice results in peri-implantation lethality (12). In addition, conditional disruption of the corresponding mcl-1 gene shows that Mcl-1 plays an important role in early lymphoid development and in the maintenance of mature lymphocytes (13).

1. Kozopas, K.M. et al. (1993) Proc Natl Acad Sci USA 90, 3516-20.
2. Yang, T. et al. (1995) J Cell Biol 128, 1173-84.
3. Sato, T. et al. (1994) Proc Natl Acad Sci USA 91, 9238-42.
4. Zhou, P. et al. (1997) Blood 89, 630-43.
5. Wang, J.M. et al. (1999) Mol Cell Biol 19, 6195-206.
6. Jourdan, M. et al. (2003) Oncogene 22, 2950-9.
7. Chao, J.R. et al. (1998) Mol Cell Biol 18, 4883-98.
8. Domina, A.M. et al. (2000) J Biol Chem 275, 21688-94.
9. Inoshita, S. et al. (2002) J Biol Chem 277, 43730-4.
10. Domina, A.M. et al. (2004) Oncogene 23, 5301-15.
11. Maurer, U. et al. (2006) Mol Cell 21, 749-60.
12. Rinkenberger, J.L. et al. (2000) Genes Dev 14, 23-7.
13. Opferman, J.T. et al. (2003) Nature 426, 671-6.

注意事项：

Storage: Mcl-1 siRNA I is supplied in RNAse-free water. Aliquot
and store at -20ºC.