| 货号 | 6620S |
| 描述 | SignalSilence® AMPKα2 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit AMPKα2 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. |
| 反应种属 | Human |
| 应用 | TFN |
| 供应商 | CST |
| 背景 | AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three distinct genes (α1, 2; β1, 2; γ1, 2, 3) (2). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia, and ischemia (1). The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop, and this phosphorylation is required for AMPK activation (3-5). AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinase and the biological significance of these phosphorylation events have yet to be elucidated (6). The β1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101, Ser108, and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization (7). Several mutations in AMPKγ subunits have been identified, most of which are located in the putative AMP/ATP binding sites (CBS or Bateman domains). Mutations at these sites lead to reduction of AMPK activity and cause glycogen accumulation in heart or skeletal muscle (1,2). Accumulating evidence indicates that AMPK not only regulates the metabolism of fatty acids and glycogen, but also modulates protein synthesis and cell growth through EF2 and TSC2/mTOR pathways, as well as blood flow via eNOS/nNOS (1). |
| 存放说明 | -20C |
| 参考文献 | Hardie, D.G. (2004) J Cell Sci 117, 5479-87. Carling, D. (2004) Trends Biochem Sci 29, 18-24. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35. Woods, A. et al. (2003) J Biol Chem 278, 28434-42. Warden, S.M. et al. (2001) Biochem J 354, 275-83. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® AMPKα2 siRNA I (+) or SignalSilence® AMPKα2 siRNA II #6630 (+), using AMPKα2 Antibody #2757 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The AMPKα2 Antibody confirms silencing of AMPKα2 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control. Western blot检测hela细胞提取物，细胞分别用100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® AMPKα2 siRNA I #6620 (+) 和 SignalSilence® AMPKα2 siRNA II (+)进行转染，所用抗体为AMPKα2 Antibody #2757 (上) 或α-Tubulin (11H10) Rabbit mAb兔单抗 #2125 (下)。AMPKα2 Antibody验证了AMPKα2基因的沉默情况，α-Tubulin (11H10) Rabbit mAb兔单抗是内参对照。 |
