SignalSilence? PERK siRNA I

• 产品编号：CST-9024S      品牌：CST       原厂货号：9024S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence® PERK siRNA I能够抑制人及猴子中PERK的表达。Cell Signaling Technology（CST）的SignalSilence^® PERK siRNA I 可以通过RNA干扰特异性抑制PERK的表达。RNA干扰是一种通过传递双链RNA分子到细胞中从而选择性沉默目的基因表达的方法。所有SignalSilence^® siRNA 产品均经过严格的内部测试并通过Western验证可有效降低目的蛋白的表达。通过三苯基分析对寡核苷酸合成进行逐个碱基的监测以保证适当的耦合效率，随后通过亲和固相提取来纯化寡核苷酸。退火的双链RNA需要通过质谱进行进一步的分析从而确定提取物的组成。为了保证批次间一致性的最大化，每一批次产品都要通过质谱与前一批次的产品进行比较。CST建议使用 100 nM SignalSilence^® PERK siRNA I 进行转染，并在48-72小时后裂解细胞。具体的转染操作步骤请根据转染试剂生产商提供的方法进行。如您在使用过程中有任何问题请随时联系我们。每瓶中含有100次转染所需用量。每次转染siRNA的终浓度为100 nM，以上用量为24孔板中转染，每孔总体积为300 μl。蛋白激酶类似的内质网激酶(PERK)是一种eIF2α激酶并且为一种定位在endoplasmic reticulum (ER) 膜上的跨膜蛋白，能够传导ER应激信号从而抑制翻译（1-3）。ER应激会提高PERK的活性，然后使eIF2α磷酸化从而增加翻译水平的降低。研究证实PERK缺失的小鼠在出生后的几周内其胰岛β细胞会出现缺陷，这也暗示着PERK介导的翻译控制在ER应激时保护分泌细胞的重要作用（4）。ER应激时PERK的活化与其胞内激酶结构域的自身磷酸化相关联（1-3）。PERK Thr980位点的磷酸化是其自身活化状态的一个标志。

1. Harding, H. et al. (1999) Nature 397, 271-274.
2. Shi, Y. et al. (1998) Mol. Cell. Biol. 18, 7499-7509.
3. Harding, H. et al. (2001) Mol. Cell 7, 1153-1163.

原厂资料：

Homology

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

Specificity / Sensitivity

SignalSilence^® PERK siRNA I inhibits human and monkey PERK expression.

Description

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

Protein kinase-like endoplasmic reticulum kinase (PERK) is an eIF2α kinase and transmembrane protein resident in the endoplasmic reticulum (ER) membrane that couples ER stress signals to translation inhibition (1-3). ER stress increases the activity of PERK, which then phosphorylates eIF2α to promote reduced translation. Research studies have demonstrated that PERK-deficient mice have defects in pancreatic β cells several weeks after birth, suggesting a role for PERK-mediated translational control in protecting secretory cells from ER stress (4). PERK activation during ER stress correlates with autophosphorylation of its cytoplasmic kinase domain (1-3). Phosphorylation of PERK at Thr980 serves as a marker for its activation status.

1. Harding, H. et al. (1999) Nature 397, 271-274.
2. Shi, Y. et al. (1998) Mol. Cell. Biol. 18, 7499-7509.
3. Harding, H. et al. (2001) Mol. Cell 7, 1153-1163.

注意事项：

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