Phospho-AMPKα (Thr172) Antibod

2531:

Western Blotting

Phospho-AMPKalpha (Thr172) Antibody detects endogenous AMPKα only when phosphorylated at threonine 172. The antibody detects both α1 and α2 isoforms of the catalytic subunit, but it does not detect the regulatory beta or gamma subunits.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr172 of human AMPKα. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of untreated and AICAR-treated C2C12 cell extracts using Phospho-AMPKα (Thr172) Antibody.

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.

• Minokoshi, Y. et al. (2002) Leptin stimulates fatty-acid oxidation by activating AMP-activated kinase. Nature 415, 339-343. Applications: Western Blotting
• Zou, M. H. et al. (2003) Activation of 5'-AMP-activated Kinase Is Mediated through c-Src and Phosphoinositide 3-Kinase Activity during Hypoxia-Reoxygenation of Bovine Aortic Endothelial Cells. The Journal of Biological Chemistry 278 (36), 34003-34010. Applications: IP Western Blotting
• Andersson, U. et al. (2004) AMP-activated Protein Kinase Plays a Role in the Control of Food Intake. The Journal of Biological Chemistry 279 (13), 12005-12008. Applications: Western Blotting
• Pilon, G. et al. (2004) Inhibition of Inducible Nitric-oxide Synthase by Activators of AMP-activated Protein Kinase. The Journal of Biological Chemistry 279 (20), 20767-20774. Applications: Western Blotting
• Shaw, R. J. et al. (2004) The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. PNAS 101 (10), 3329-3335. Applications: Western Blotting
• Gonzalez, A.A. et al. (2004) Am J Physiol Endocrinol Metab 287, E1032-7. Applications: Western Blotting

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• 9996 Oligomycin

For Research Use Only. Not For Use In Diagnostic Procedures.