SignalSilence? PKA C-α siRNA II

• 产品编号：CST-6574S      品牌：CST       原厂货号：6574S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

SignalSilence PKA C-α siRNA II会抑制人、小鼠和大鼠的PKA C-α表达。Cell Signaling Technology (CST)公司生产的PKA C-α siRNA II使得研究者可以使用RNA干扰技术特异性的抑制PKA C-α的表达。这种技术通过将双链RNA分子送入细胞而选择性的沉默某个基因的表达。CST的siRNA产品已经在内部经过严格测试，经western分析确能减少目标蛋白的表达。CST公司推荐您在细胞裂解前48-72小时用100nM PKA C-α siRNA进行转染。转染的流程请遵循转染试剂制造商提供的操作流程。使用中如有任何问题，请与CST联系。第二信使环磷酸腺苷（cAMP）在哺乳动物细胞中激活cAMP依赖的蛋白激酶（PKA或者cAPK），控制如基因转录，离子转运和蛋白磷酸化等许多细胞内机制（1）。失活的PKA的是一个由调节亚基（R）和催化亚基（C）组成的异二聚体。在失活状态时，R亚基的假底物序列阻止了C亚基的活性中心。已识别出三个催化亚基的亚型（C-α, C-β, 和C-γ）和两个调节亚基家族（RI和RII），其分别具有不同的cAMP结合特性。两个R家族又进一步存在两种亚型，α和β (RI-α, RI-β, RII-α和RII-β)。R亚基与cAMP结合后，自身抑制性接触被解除，有活性的C亚基单体被释放。PKA与Akt（PKB）和PKC共有相同的底物特异性，以一个在3位的与酪氨酸和苏氨酸残疾磷酸化相关的精氨酸为特点（2）。具有这些相同结构序列的底物已经被证实能被PKA磷酸化，它们包括Bad (Ser155), CREB (Ser133), 和GSK-3 (GSK-3α Ser21和GSK-3β Ser9)(3-5)。另外，同时下调PKA C-α和-β可以阻止cAMP介导的Raf（Ser43和Ser259）磷酸化（6）。自身磷酸化和被PDK-1磷酸化是C亚基Thr197磷酸化的两个已知的机制（7）。

1. Montminy, M. (1997) Annu. Rev. Biochem. 66, 807-822.
2. Dell'Acqua, M.L. and Scott, J.D. (1997) J. Biol. Chem. 272, 12881-12884.
3. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
4. Gonzalez, G.A. and Montminy, M.R. (1989) Cell 59, 675-680.
5. Fang, X. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 11960-11965.
6. Dumaz, N. and Marais, R. (2003) J. Biol. Chem. 278, 29819 -29823.
7. Moore, M.J. et al. (2002) J. Biol. Chem. 277, 47878-47884.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Mouse, Rat

Specificity / Sensitivity

SignalSilence® PKA C-α siRNA II will inhibit human, mouse and rat PKA C-α exoression.

Description

SignalSilence® PKA C-α siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PKA C-α 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 are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Directions for Use

CST recommends transfection with 100 nM PKA C-α siRNA 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

The second messenger cyclic AMP (cAMP) activates cAMP-dependent protein kinase (PKA or cAPK) in mammalian cells and controls many cellular mechanisms such as gene transcription, ion transport, and protein phosphorylation (1). Inactive PKA is a heterotetramer composed of a regulatory subunit (R) dimer and a catalytic subunit (C) dimer. In this inactive state, the pseudosubstrate sequences on the R subunits block the active sites on the C subunits. Three C subunit isoforms (C-α, C-β, and C-γ) and two families of regulatory subunits (RI and RII) with distinct cAMP binding properties have been identified. The two R families exist in two isoforms, α and β (RI-α, RI-β, RII-α, and RII-β). Upon binding of cAMP to the R subunits, the autoinhibitory contact is eased and active monomeric C subunits are released. PKA shares substrate specificity with Akt (PKB) and PKC, which are characterized by an arginine at position -3 relative to the phosphorylated serine or threonine residue (2). Substrates that present this consensus sequence and have been shown to be phosphorylated by PKA are Bad (Ser155), CREB (Ser133), and GSK-3 (GSK-3α Ser21 and GSK-3β Ser9) (3-5). In addition, combined knock-down of PKA C-α and -β blocks cAMP-mediated phosphorylation of Raf (Ser43 and Ser259) (6). Autophosphorylation and phosphorylation by PDK-1 are two known mechanisms responsible for phosphorylation of the C subunit at Thr197 (7).

1. Montminy, M. (1997) Annu. Rev. Biochem. 66, 807-822.
2. Dell'Acqua, M.L. and Scott, J.D. (1997) J. Biol. Chem. 272, 12881-12884.
3. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
4. Gonzalez, G.A. and Montminy, M.R. (1989) Cell 59, 675-680.
5. Fang, X. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 11960-11965.
6. Dumaz, N. and Marais, R. (2003) J. Biol. Chem. 278, 29819 -29823.
7. Moore, M.J. et al. (2002) J. Biol. Chem. 277, 47878-47884.

注意事项：

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