Biolegend, 600651, Purified anti-AMPKα Phospho (Thr172) Antibody, 25μg

AMP-activated protein kinase alpha (AMPKα) is one subunit of the αβγ heterotrimeric protein complex AMPK. It is a key regulator of metabolism and energy homeostasis in eukaryotes through induction of catabolic pathways and deactivation of anabolic. As an inhibitor of mammalian target of rapamycin complex 1 (mTORC1) pathway, AMPKα is involved in the regulation of cellular growth, cell cycle progression, autophagy, and may play dual roles as both tumor suppressor and promotor. AMPKα is activated via phosphorylation of Threonine residue 172 (Thr172) within the activation loop of the α subunit and is promoted by the allosteric binding of AMP and/or ADP to AMPK’s γ subunit. This phosphorylation is mediated by the serine/threonine kinase liver kinase B1 (LKB1) in conjunction with accessory subunits STRAD and MO25 and also by calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK2 aka CAMKKβ). Activation is induced by increases in cellular AMP:ATP and ADP:ATP ratios  following a decline in ATP levels as well as by cellular stress, DNA damage, glucose starvation, and fluxes in calcium and nutrient levels. Localization of AMPKα Thr172 at the spindle poles suggests a novel role for AMPK in mitotic spindle orientation. AMPKα is a key therapeutic target in the treatment of metabolic disorders and an emerging potential treatment target for cancer.
25μg
Verified Reactivity: Human, Mouse
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Synthetic peptide of human AMPK alpha phosphorylated at Thr172
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C.
Application: WB - Quality tested ICC, ICFC, IHC-P - Verified
Recommended Usage: Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration of 5.0 μg/mL is recommended. For flow cytometric staining using our True-Phos™ Perm Buffer, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: This clone was tested for ICC using serum-starved untreated HEK293 cells (negative control) and HEK293 cells treated with 5.0 µM oligomycin for 30 minutes (positive control). Staining was expected to localize to the nucleus and cytoplasm. Three fix/perm methods were used: methanol fixation or 4% PFA fixation followed by permeabilization with either methanol or Triton X-100. Both PFA fixation followed by methanol permeabilization and PFA fixation followed by Triton X-100 permeabilization produced a strong signal with the expected localization. Methanol-only fixation only produced a faint signal.
RRID: AB_2904438 (BioLegend Cat. No. 600651) AB_2904438 (BioLegend Cat. No. 600652)
Structure: AMPKα is a 552 amino acid protein with a predicted molecular weight of 63 kD.
Distribution: Heart, Skeletal muscle, Kidney / Nucleus and cytoplasm
Function: Serine/threonine-protein kinase, transcription regulation, Wnt signaling, metabolism
Ligand/Receptor: ATP-binding, magnesium, Metal-binding, Nucelotide-binding
Biology Area: Cell Biology, Signal Transduction
Molecular Family: Phospho-Proteins, Protein Kinases/Phosphatase
Antigen References: Mihaylova MM, et al. 2011. Nat Cell Biol 13:1016-23. Stein SC, et al. 2000. Biochem J. 345:437-43. Thaiparambil JT, et al. 2012. Mol Cell Biol. 16:3203-17. Vara-Ciruelos D, et al. 2019. Open Biol. 9:190099. Willows R, et al. 2017. Biochem J. 474:3059-3073.
Gene ID: 55635562
UniProt: View information about AMPKalpha Phospho Thr172 on UniProt.org
Clone: A20017A
Regulatory Status: RUO
Other Names: PRKAA1, PRKAA2, AMPKa1, AMPKa2, AMPK, ACACA Kinase, Tau-Protein Kinase PRKAA1, Tau-Protein Kinase PRKAA2, Protein kinase AMP-Activated, Alpha 1 Catalytic Subunit, HMGCR Kinase
Isotype: Mouse IgG1, κ