Phospho-AMPKα1 (Ser485) (45F5)

2537:

Western Blotting

Phospho-AMPKα1 (Ser485) (45F5) Rabbit mAb detects endogenous levels of AMPKα1 only when phosphorylated at Ser485. The antibody does not cross-react with phosphorylated AMPKα2 or other related proteins.

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser485 of human AMPKα1.

Western Blotting

Western blot analysis of HEK293 cell lysates treated with λ phosphatase or AICAR, using Phospho-AMPKα1 (Ser485) (45F5) Rabbit mAb (upper) and AMPKα Antibody #2532 (lower).

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.

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• 2603 AMPKα (23A3) Rabbit mAb
• 2793 AMPKα (F6) Mouse mAb
• 2795 AMPKα1 Antibody
• 2532 AMPKα Antibody
• 4188 Phospho-AMPKα (Thr172) (D79.5E) Rabbit mAb
• 2535 Phospho-AMPKα (Thr172) (40H9) Rabbit mAb
• 2531 Phospho-AMPKα (Thr172) Antibody
• 4184 Phospho-AMPKα1 (Ser485) Antibody
• 4185 Phospho-AMPKα1 (Ser485)/AMPKα2 (Ser491) Antibody
• 2757 AMPKα2 Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9944 AICAR
• 9996 Oligomycin

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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