AMPK Subunit Antibody Sampler Kit

• 产品编号：CST-9839S      品牌：Cell Signaling Technology       原厂货号：9839S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

每一个AMPK Subunit Antibody Sampler Kit中的抗体都检测特定的内源性AMPK蛋白水平。抗体不与相关的AMPK亚基交叉反应。通过类人AMPKα碳端Leu519周围残基，类人AMPKα2 Ser500位点周围残基，类人AMPKγ1氨基端残基，类人AMPKγ2 Ser60位点周围残基，类人AMPKβ2和AMPKγ3氨基酸序列的合成肽免疫动物 得到多克隆抗体。抗体的纯化采用蛋白A和肽段亲和层析的方法。通过类人AMPKβ1 val176周围残基序列的合成肽免疫动物得到单克隆抗体。AMPK Subunit Antibody Sampler Kit为研究AMPK各个亚基在细胞能量平衡中发挥的作用提供了一种经济有效的方法。试剂盒中的每种抗体都至少能够满足四次Western blot实验所需的一抗和二抗。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

Each of the antibodies in the AMPK Subunit Antibody Sampler Kit detects endogenous levels of the specified AMPK protein. Antibodies do not cross-react with related AMPK subunit proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu519 near the carboxy terminus of human AMPKα1,corresponding to residues surrounding Ser500 of human AMPKα2, near the amino terminus of human AMPKγ1, surrounding Ser60 of human AMPKγ2, and corresponding to the sequences of human AMPKβ2 and AMPKγ3. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Val176 of human AMPKβ1.

Description

The AMPK Subunit Antibody Sampler Kit provides an economical means to investigate the role played by all AMPK subunits in cellular energy homeostasis. The kit contains enough primary and secondary antibodies to perform four Western blots with each antibody.

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 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies.