Anti-phospho-AMPK alpha-1 Antibody, (Thr479)

5′-AMP-activated protein kinase catalytic subunit alpha-1 (EC 2.7.11.1; EC 2.7.11.26; EC 2.7.11.27; EC 2.7.11.31; UniProt Q13131; also known as ACACA kinase, Acetyl-CoA carboxylase kinase, AMPK subunit alpha-1, AMPKa1, HMGCR kinase, Hydroxymethylglutaryl-CoA reductase kinase, Tau-protein kinase PRKAA1) is encoded by the PRKAA1 (also known as AMPK1) gene (Gene ID 5562) in human. The yeast sucrose non-fermenting 1 (SNF1) and the AMP-activated protein kinase (AMPK) in drosphila and mammals are orthologous heterotrimeric serine-threonine kinases, each consisting of a catalytic α subunit and regulatory β and γ subunits, that sense decreased cellular energy levels when the AMP to ATP ratio rises due to a high rate of metabolism or decreased nutrient availability. AMP binding to the γ subunit results in an enhanced phosphorylation of the catalytic α subunit on an activation loop threonine and AMPK allosteric activation. Activated AMPK phosphorylates downstream targets, leading to energy conservation and inhibition of ATP consumption. For example, the high rate of ATP use in contracting muscle causes AMPK-mediated phosphorylation/inactivation of acetyl-CoA carboxylase (ACC), thereby increasing fatty acid oxidation and restoring cellular energy. AMPK functions as a positive regulator of autophagy by phosphorylating the protein kinase ULK1 that initiates autophagy as well as by inhibiting the mammalian TOR (mTOR) pathway activity via phosphorylating mTOR-binding partner raptor. In mammals, there exist two different α (encoded by the PRKAA1 and PRKAA2 gene), two different β (encoded by the PRKAB1 and PRKAB2 gene), and three different γ (encoded by the PRKAG1, PRKAG2, and PRKAG3 gene) subunits.

The Thr479 numbering is based on the rat AMPK alpha-1 sequence shown in Fig 2A of Suzuki, T., et al. (2013). Mol. Cell. 50(3):407-419, which was cited for as Thr479 phosphorylation in subsequent publications (Coughlan, K.A., et al. (2015). PLoS One. 10(5):e0127388; Hawley, S.A., et al. (2014). Biochem. J. 459(2):275-287). The target threonine corresponds to Thr490 of the human/mouse/rat AMPK alpha-1 sequences published by UniProt (Q13131-1/Q5EG47/P54645) or Thr505 of human spliced isoform 2 (UniProt.Q13131-2). The target site sequence is not well conserved between AMPK alpha-1 (-SGTA(pT)PQR-) and AMPK alpha-2 (-SGSS(pT)PQR-).

KLH-conjugated linear peptide corresponding to the AMPK alpha-1 phosphorylation site sequence containing the phosphorylated target threonine.

Detect AMPK threonine 479 phosphorylation using this rabbit polyclonal Anti-phospho-AMPK alpha-1 (Thr479), Cat. No. ABS981, validated for use in Dot Blot and Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A 1:500 dilution from a representative lot detected AMPK alpha-1 phosphorylation induction upon 30-min serum stimulation of 6-hr serum-starved HEK293 cells (Courtesy of Bruce Kemp, Ph.D., St. Vincent′s Institute of Medical Research in Melbourne).

Western Blotting Analysis: A 1:1000 dilution from a representative lot detected GSK3beta-catalyzed phosphorylation of E. coli expressed, catalytically inactive AMPK alpha-1 recombinant protein (Courtesy of Bruce Kemp, Ph.D., St. Vincent′s Institute of Medical Research in Melbourne).

Evaluated by Peptide Dot Blot.

Dot Blot Analysis: A 1:1,000 dilution of this antibody detected 20-0.156 ng of immunogen peptide, but not the corresponding non-phosphorylated peptide.

~63 kDa observed. 64.01/65.52 kDa (human isoform 1/2) and 63.93/63.97 kDa (mouse/rat) calculate. Uncharacterized bands may be observed in some lysate(s).

Affinity purified.

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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