Cell Signaling Technology

Product Pathways - Glucose Metabolism

Phospho-AMPKα (Thr172) (D79.5E) Rabbit mAb #4188

Applications Reactivity MW (kDa) Source Isotype
W H M R Dr (B) 62 Rabbit IgG

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

Specificity / Sensitivity

Phospho-AMPKα (Thr172) (D79.5E) Rabbit mAb detects endogenous AMPKα only when phosphorylated at Thr172. This antibody detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Thr172 of human AMPKα.

Western Blotting

Western Blotting

Western analysis of extracts from SK-N-MC cells, untreated or AICAR-treated, using Phospho-AMPKα (Thr172) (D79.5E) Rabbit mAb (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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Product Pathways

Drug Discovery Tools

Featured Technologies

Protein Classes