PathScan? Phospho-AMPKα (Thr172) Sandwich ELISA Antibody Pair

• 产品编号：CST-7955S      品牌：CST       原厂货号：7955S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
• 应用分类：

其它组分：

描述：

抗体对的特异性和灵敏性请参考相关PathScan® Sandwich ELISA Kit。内控检测证明，抗体对用于检测相应物种的对应蛋白，但同时也可与其它物种的同源蛋白交互反应。CS''s PathScan® Phospho-AMPKα (Thr172) Sandwich ELISA Antibody Pair是PathScan® Phospho-AMPKα-(Thr172) Sandwich ELISA Kit #7959产品的更经济实惠的替代型产品。产品包含有捕获抗体和检测抗体(100X原液)，以及抗鼠IgG，辣根过氧化物酶耦联的抗体(1000X原液)。所含试剂可以做四个96孔的ELISA。AMPKα Rabbit Capture Antibody首先在96孔板中于PBS中孵育过夜，使之紧实地结合到孔中。捕获抗体结合后，加入细胞裂解液，然后再加入Phospho-AMPKα (Thr172) Mouse Detection Antibody和Anti-Mouse IgG, HRP-linked Antibody。然后加入辣根过氧化物酶的底物TMB使之显色。显色的强度与磷酸化AMPKα (Thr172)蛋白的浓度成正比。试剂盒中的抗体都是定制的。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：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST''s PathScan® Phospho-AMPKα (Thr172) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-AMPKα-(Thr172) Sandwich ELISA Kit #7959. Capture and Detection antibodies (100X stocks) and Anti-Mouse IgG, HRP-linked Antibody (1000X stock) are supplied. Sufficient reagents are provided for 4 x 96 well ELISAs. The AMPKα Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Phospho-AMPKα (Thr172) Mouse Detection Antibody and Anti-Mouse IgG, HRP-linked Antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-AMPKα (Thr172) protein. 
 Antibodies in kit are custom formulations specific to kit.

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: Capture and Detection Antibodies are stored at 4ºC.
HRP-Linked Streptavidin is stored at -20ºC.