Cell Signaling Technology

Product Pathways - Glucose Metabolism

Phospho-AMPKα1 (Ser485) Antibody #4184

Applications Reactivity MW (kDa) Source
W H M R Mk (C) 62 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-AMPKα1 (Ser485) Antibody detects endogenous levels of AMPKα1 only when phosphorylated at serine 485. The antibody does not cross-react with phosphorylated AMPKα2 or other related proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser485 of human AMPKα1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEK293 and C6 cells, λ phosphatase-treated or oligomycin-treated, using Phospho-AMPKα1 (Ser485) Antibody (upper) or AMPKα Antibody #2532 (lower).

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, 5479-5487.
  2. Carling, D. (2004) Trends Biochem. Sci. 29, 18-24.
  3. Hawley, S.A. et al. (1996) J. Biol. Chem. 271, 27879-27887.
  4. Lizcano, J.M. et al. (2004) EMBO J. 23, 833-843.
  5. Shaw, R. et al. (2004) Proc. Natl. Acad. Sci. USA 101, 3329-3335.
  6. Woods, A. et al. (2003) J. Biol. Chem. 278, 28434-28442.
  7. Warden, S.M. et al. (2001) Biochem. J. 354, 275-283.

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