AMP Activated Protein Kinase alpha, phosphorylated (Thr172) (AMPKa)

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. AMPK is a heterotrimeric complex composed of a catalytic alpha subunit and regulatory beta and gamma subunits, each of which is encoded by two or three distinct genes (a1, 2; b1, 2; g1, 2, 3). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia and ischemia. The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPK-alpha at Thr172 in the activation loop and this phosphorylation is required for AMPK activation. AMPK-alpha is also phosphorylated at Thr258 and Ser485 (for a1; Ser491 for a2). The upstream kinase and the biological significance of these phosphorylation events have yet to be elucidated. The beta1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101, Ser108 and Ser182. Phosphorylation at Ser108 of the beta1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization. Several mutations in AMPK-gamma 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. 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.\n\nApplications: \nSuitable for use in ELISA and Western Blot. Other applications have not been tested.\n\nRecommended Dilutions: \nWestern Blot: 1:1000. Band at ~62kD.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability: \nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

A1475-05

Size

100ul

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.

P Type

Pab

Purity

Purified by Protein A and peptide affinity chromatography.

Isotype

IgG

References

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.

Additional Info

Recognizes endogenous human AMPK-alpha at ~62kD only when activated by phosphorylation at Thr172. Detects both alpha-1 and alpha-2 isoforms of the catalytic subunit. Does not detect the regulatory beta or gamma subunits. Species Crossreactivity: mouse, rat and monkey.