CST, 50081T, Phospho-AMPK alpha (Thr172) (D4D6D) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying AMPK-alpha1 (Thr183) phosphate/AMPK-alpha2 (Thr172) phosphate. Validated for WB,WB,IP,IHC. Available in 2 sizes. Highly specific and rigorously validated in-house, Phospho-AMPK alpha (Thr172) (D4D6D) Rabbit Monoclonal Antibody (CST #50081) is ready to ship. Product Usage Information Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:50 Immunohistochemistry (Paraffin): 1:200 - 1:800 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin) Specificity / Sensitivity Phospho-AMPK alpha (Thr172) (D4D6D) Rabbit Monoclonal Antibody detects endogenous AMPKα only when phosphorylated at threonine 172. This antibody detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr172 of human AMPKα protein. 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 AMPK activation, 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). Alternate Names 5'-AMP-activated protein kinase catalytic subunit alpha-1; 5'-AMP-activated protein kinase catalytic subunit alpha-2; 5'-AMP-activated protein kinase, catalytic alpha-1 chain; 5'-AMP-activated protein kinase, catalytic alpha-2 chain; AAPK1; AAPK2; ACACA kinase; Acetyl-CoA carboxylase kinase; AMP -activate kinase alpha 1 subunit; AMP-activated protein kinase; AMP-activated protein kinase alpha-2 subunit variant 2; AMP-activated protein kinase alpha-2 subunit variant 3; AMP-activated protein kinase, catalytic, alpha-1; AMPK; AMPK alpha 1; AMPK alpha 2; AMPK subunit alpha-1; AMPK subunit alpha-2; AMPK-alpha-2 chain; AMPK1; AMPK2; AMPKa1; AMPKA2; HMGCR kinase; Hydroxymethylglutaryl-CoA reductase kinase; MGC33776; MGC57364; PRKAA; PRKAA1; PRKAA2; protein kinase AMP-activated catalytic subunit alpha 1; protein kinase AMP-activated catalytic subunit alpha 2; protein kinase, AMP-activated, alpha 1 catalytic subunit; protein kinase, AMP-activated, alpha 2 catalytic subunit; Tau-protein kinase PRKAA1 Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 62 Source/Isotype: Rabbit IgG