SignalSilence? p38α MAPK siRNA I

• 产品编号：CST-6269S      品牌：CST       原厂货号：6269S
• 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^® p38α MAPK siRNA I可以帮助研究者通过RNA干扰特异性地抑制p38α MAPK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 p38α MAPK 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。p38 MAP kinase (MAPK),也被称为RK (1)或CSBP (2)，是酵母中HOG激酶在哺乳动物中的同源蛋白，在细胞因子和应急诱导的信号转导过程中起重要作用。(1-4)。已经发现了4种亚型的p38 MAPK，分别是p38α, β, γ（也被称为Erk6或 SAPK3）以及δ (也被称为 SAPK4)。与SAPK/JNK通路类似，p38 MAPK可以被多种细胞压力所激活，包括渗透压休克，炎症因子，脂多糖（LPS），紫外线照射和生长因子（1-5）。MKK3, MKK6和SEK能够使p38 MAPK的Thr180 和Tyr182位点磷酸化，从而激活p38 MAPK。激活的p38 MAPK能够磷酸化并激活MAPKAP kinase 2 (3)，进而最终磷酸化ATF-2 (5), Max (6)和MEF2等转录因子(5-8)。SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole)是p38 MAPK的选择性抑制剂。该复合物通过抑制p38 MAPK及后续HSP27的磷酸化，抑制MAPKAPK-2的激活（9）。SB203580能够结合到p38 MAPK与ATP结合的位置从而抑制p38 MAPK的催化活性，但不会通过上游激酶抑制p38 MAPK的磷酸化（10）。

1. Rouse, J. et al. (1994) Cell 78, 1027-37.
2. Han, J. et al. (1994) Science 265, 808-11.
3. Lee, J.C. et al. (1994) Nature 372, 739-46.
4. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
5. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
7. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
8. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

原厂资料：

Description

SignalSilence® p38α MAPK siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit p38α MAPK 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 p38α MAPK 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

p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). 
 SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).

1. Rouse, J. et al. (1994) Cell 78, 1027-37.
2. Han, J. et al. (1994) Science 265, 808-11.
3. Lee, J.C. et al. (1994) Nature 372, 739-46.
4. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
5. Raingeaud, J. et al. (1995) J Biol Chem 270, 7420-6.
6. Zervos, A.S. et al. (1995) Proc Natl Acad Sci U S A 92, 10531-4.
7. Zhao, M. et al. (1999) Mol Cell Biol 19, 21-30.
8. Yang, S.H. et al. (1999) Mol Cell Biol 19, 4028-38.
9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

注意事项：

Storage: p38α MAPK siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.