AMPK Control Cell Extracts

• 产品编号：CST-9158S      品牌：CST       原厂货号：9158S
• 产品分类：蛋白质表达与纯化 > 蛋白质样本制备 > 细胞组织粗提物与裂解物
• 应用分类：

描述：

Untreated AMPK Control Cell Extracts：用培养于含血清培养液的C2C12细胞提取物作为阴性对照。 
Treated AMPK Control Cell Extracts: 用血清饥饿的C2C12细胞提取物作为阳性对照。用作对照，CST建议两种对照液的推荐使用量均为20 µl。使用前请将样品煮沸3-5分钟。腺苷酸激活的蛋白激酶（AMPK）是一种在酵母，植物和动物界高度保守的蛋白激酶，它在能量平衡中发挥关键性的作用(1)。AMPK是异源三聚体复合物，由α催化亚基和β、γ两个调节亚基组成，每个亚基都由两到三个不同的基因编码(α1, 2; β1, 2; γ1, 2, 3)(2)。细胞或者环境因素的刺激，如热激，缺氧，缺血等，导致AMP/ATP浓度比的升高，AMP/ATP浓度比的升高可以激活AMPK。肿瘤抑制因子LKB1，与辅助蛋白STRAD和MO25相结合，在AMPK的α亚基活化环的172位苏氨酸上对其磷酸化，磷酸化对AMPK的激活是必需的(3-5)。AMPKα同样也可以在Thr258和Ser485(α1; α2是Ser491)位点磷酸化。此过程的上游激酶和这些位点磷酸化的生物学意义尚不明晰(6)。β1亚基有一些翻译后的修饰，包括豆蒄酰化修饰，以及在多个位点的磷酸化修饰(包括Ser24/25, Ser96, Ser101和Ser182)(6,7)。β1亚基Ser108的磷酸化可能对AMPK的激活来说是必需的，而Ser24/25和Ser182的磷酸化可能影响到AMPK的定位(7)。越来越多的证据表明，AMPK不仅调控机体的糖脂代谢，同时也通过EF2和TSC2/mTOR通路调控细胞生长和蛋白质的合成，以及通过eNOS/nNOS系统来调节血流量(1)。

1. Hardie, D.G. (2004) J Cell Sci 117, 5479-87.
2. Carling, D. (2004) Trends Biochem Sci 29, 18-24.
3. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87.
4. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
5. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35.
6. Woods, A. et al. (2003) J Biol Chem 278, 28434-42.
7. Warden, S.M. et al. (2001) Biochem J 354, 275-83.

原厂资料：

Description

Untreated AMPK Control Cell Extracts: Total cell extracts from C2C12 cells, prepared with serum, serve as a negative control. 
 Treated AMPK Control Cell Extracts: Total cell extracts from C2C12 cells, prepared by serum starvation, serve as a positive control.

Applications

As controls, CST recommends using 20 µl of untreated and treated AMPK Control Cell Extract. Boil sample for 3-5 minutes before use.

Background

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).

1. Hardie, D.G. (2004) J Cell Sci 117, 5479-87.
2. Carling, D. (2004) Trends Biochem Sci 29, 18-24.
3. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87.
4. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
5. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35.
6. Woods, A. et al. (2003) J Biol Chem 278, 28434-42.
7. Warden, S.M. et al. (2001) Biochem J 354, 275-83.

注意事项：

Storage: Supplied in SDS Sample Buffer: 62.5 mM Tris-
HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM
DTT, 0.01% w/v bromophenol blue or phenol red. Store at
-20°C, or at -80°C for long-term storage.