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Purified anti-AMPKα Phospho (Thr172) Antibody

Pricing & Availability
Clone
A20017A (See other available formats)
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
Ave. Rating
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Product Citations
publications
A20017A_PURE_Aquaporin-4_Antibody_1_120221
Whole cell extracts (15 µg total protein) from serum-starved HEK293 cells treated with (+) or without (-) 5.0 µM oligomycin for 30 minutes (+) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-AMPKα Phospho (Thr172) antibody (clone A20017A) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-β-actin Antibody (Cat. No. 643808) was used as a loading control at a 1:10000 dilution (lower). Lane M: Molecular weight marker
  • A20017A_PURE_Aquaporin-4_Antibody_1_120221
    Whole cell extracts (15 µg total protein) from serum-starved HEK293 cells treated with (+) or without (-) 5.0 µM oligomycin for 30 minutes (+) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-AMPKα Phospho (Thr172) antibody (clone A20017A) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-β-actin Antibody (Cat. No. 643808) was used as a loading control at a 1:10000 dilution (lower). Lane M: Molecular weight marker
  • A20017A_PURE_Aquaporin-4_Antibody_2_120221
    Whole cell extracts (15 µg total protein) from untreated C2C12 cells (-) and C2C12 cells treated with 5.0 µM oligomycin for 30 minutes (+) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-AMPKα Phospho (Thr172) antibody (clone A20017A) for 2 hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker
  • A20017A_PURE_Aquaporin-4_Antibody_3_120221
    Serum-starved untreated HEK293 cells (negative control, panel A) and HEK293 cells treated with 5.0 µM oligomycin for 30 minutes (positive control, panel B) were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with purified anti-AMPKα Phospho (Thr172) antibody (clone A20017A) overnight at 4°C followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405326) at 5.0 µg/mL. Nuclei were counterstained with DAPI and the images were captured with a 60X objective.
  • A20017A_PURE_Aquaporin-4_Antibody_4_120221
    HEK293 cells were either untreated (negative control, open histogram), or treated with Oligomycin (positive control, filled histogram). The cells were fixed and permeabilized using the True-Phos™ Perm Buffer Set (Cat. No. 425401) and intracellularly stained with Purified anti-AMPKα Phospho (Thr172) (clone A20017A) or Purified Mouse IgG1, κ Isotype Control (open histogram, dashed line) (representative histogram for either untreated or treated cells) (Cat. No. 401402) followed by Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322).
  • A20017A_PURE_Aquaporin-4_Antibody_5_120221
    HC staining of Purified anti-AMPKα Phospho (Thr172) on formalin-fixed paraffin-embedded human colon was performed following antigen retrieval using Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422703). Tissue sections were incubated with Alexa Fluor® 647 Goat anti-mouse only (Cat. No. 405322) (panel A) or with Purified anti-AMPKα Phospho (Thr172) (clone A20017A) followed by Alexa Fluor® 647 Goat anti-mouse (Cat. No. 405322) (panel B). Nuclei were counterstained with DAPI (Cat. No. 422801). Scale bar: 50 µm
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600651 25 µg $141
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600652 100 µg $329
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Description

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.

Product Details
Technical data sheet

Product Details

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)

Antigen Details

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
  1. Mihaylova MM, et al. 2011. Nat Cell Biol 13:1016-23.
  2. Stein SC, et al. 2000. Biochem J. 345:437-43.
  3. Thaiparambil JT, et al. 2012. Mol Cell Biol. 16:3203-17.
  4. Vara-Ciruelos D, et al. 2019. Open Biol. 9:190099.
  5. Willows R, et al. 2017. Biochem J. 474:3059-3073.

 

 

Gene ID
5563 View all products for this Gene ID 5562 View all products for this Gene ID
UniProt
View information about AMPKalpha Phospho Thr172 on UniProt.org

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Go To Top Version: 3    Revision Date: 06/27/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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