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Purified anti-AMPKα Antibody

Pricing & Availability
Clone
5C9-5B10-6D3 (See other available formats)
Regulatory Status
RUO
Other Names
PRKAA1, AMPKa1, AMPK, ACACA Kinase, Tau-Protein Kinase PRKAA1, Protein kinase AMP-Activated, Alpha 1 Catalytic Subunit , HMGCR Kinase
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
5C9-5B10-6D3_PURE_anti-AMPKalpaha_Antibody_121521
Whole cell extracts (15 µg total protein) from A-431, HeLa, HEK293, and NIH-3T3 were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.5 μg/mL purified anti-AMPKα antibody (clone 5C9-5B10-6D3) overnight at 4°C. 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 1:50000 dilution (lower). Lane M: Molecular weight marker
  • 5C9-5B10-6D3_PURE_anti-AMPKalpaha_Antibody_121521
    Whole cell extracts (15 µg total protein) from A-431, HeLa, HEK293, and NIH-3T3 were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.5 μg/mL purified anti-AMPKα antibody (clone 5C9-5B10-6D3) overnight at 4°C. 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 1:50000 dilution (lower). Lane M: Molecular weight marker
  • 5C9-5B10-6D3_PURE_anti-AMPKalpaha_Antibody_2_121521
    HeLa cells were fixed with ice cold methanol for 10 minutes and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL of purified mouse IgG1, κ isotype control (Cat. No. 400102) (panel A) or with 1.0 µg/mL of purified anti-AMPKα antibody (clone 5C9-5B10-6D3) (panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) at 2.5 µg/mL. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • 5C9-5B10-6D3_PURE_anti-AMPKalpaha_Antibody_3_121521
    IHC staining of anti-AMPKα antibody (clone 5C9-5B10-6D3) on formalin-fixed paraffin-embedded human colon tissue. Following antigen retrieval using Sodium Citrate H.I.E.R. (Cat. No. 928502), the tissue was incubated with 5.0 µg/mL of the primary antibody overnight at 4°C. BioLegend’s Ultra Streptavidin HRP Kit (Multi-Species, DAB) (Cat. No. 929501) was used for detection followed by hematoxylin counterstaining, according to the protocol provided. The image was captured with a 40X objective. Scale bar: 50 μm
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600551 25 µg $118
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600552 100 µg $293
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Description

AMP-activated protein kinase alpha (AMPKα) is a subunit of the αβγ heterotrimeric AMPK protein complex. It is a key regulator of metabolism and energy homeostasis in eukaryotes through induction of catabolic pathways and deactivation of anabolic pathways. 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 a dual role 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 2 (CAMKK2 aka CAMKKβ). Activation is induced by increases in the 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
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
IHC-P, ICC - 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 0.25 - 0.5µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

 

Application Notes

This clone was also tested for ICC using 4% PFA fixed HeLa cells, Triton X-100 or MeOH permeabilization, and produced mainly nuclear staining.

RRID
AB_2910471 (BioLegend Cat. No. 600551)
AB_2910471 (BioLegend Cat. No. 600552)

Antigen Details

Structure
AMPKα is a 559 amino acid protein with a predicted molecular weight of 64 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 S, 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
5562 View all products for this Gene ID
UniProt
View information about AMPKalpha on UniProt.org

Other Formats

View All AMPKα Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-AMPKα 5C9-5B10-6D3 WB,IHC-P,ICC
Go To Top Version: 1    Revision Date: 12/15/2021

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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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