Overview

  • Product nameAnti-AMPK beta 1 antibody [Y367]
    See all AMPK beta 1 primary antibodies
  • Description
    Rabbit monoclonal [Y367] to AMPK beta 1
  • SpecificityThis antibody is specific for human AMPK beta 1.
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human AMPK beta 1 aa 150-250. The exact sequence is proprietary.
    Database link: Q9Y478

  • Positive control
    • NIH 3T3, Hela, A431 and PC12, MCF-7 cell lysates. Human lung carcinoma tissue.
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Applications

Our Abpromise guarantee covers the use of ab32112 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 30 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/80.
ICC/IF 1/500.

Target

  • FunctionNon-catalytic subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Beta non-catalytic subunit acts as a scaffold on which the AMPK complex assembles, via its C-terminus that bridges alpha (PRKAA1 or PRKAA2) and gamma subunits (PRKAG1, PRKAG2 or PRKAG3).
  • Sequence similaritiesBelongs to the 5'-AMP-activated protein kinase beta subunit family.
  • DomainThe glycogen-binding domain may target AMPK to glycogen so that other factors like glycogen-bound debranching enzyme or protein phosphatases can directly affect AMPK activity.
  • Post-translational
    modifications
    Phosphorylated when associated with the catalytic subunit (PRKAA1 or PRKAA2). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK.
  • Information by UniProt
  • Database links
  • Alternative names
    • 1300015D22Rik antibody
    • 5''-AMP-activated protein kinase subunit beta-1 antibody
    • AAKB1_HUMAN antibody
    • AMP-ACTIVATED PROTEIN KINASE, NONCATALYTIC, BETA-1 antibody
    • AMP-activated, noncatalytic, beta-1 antibody
    • AMPK antibody
    • AMPK beta 1 chain antibody
    • AMPK subunit beta-1 antibody
    • AMPK-BETA-1 antibody
    • AMPKb antibody
    • AU021155 antibody
    • E430008F22 antibody
    • HAMPKb antibody
    • MGC17785 antibody
    • PRKAB1 antibody
    • Protein kinase AMP activated non catalytic subunit beta 1 antibody
    see all

Anti-AMPK beta 1 antibody [Y367] images



  • Predicted band size : 30 kDa
    Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: AMPK beta 1 knockout HAP1 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: A431 cell lysate
    Lanes 1 - 4: Merged signal (red and green). Green - ab32112 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab32112 was shown to specifically react with AMPK beta 1 when AMPK beta 1 knockout samples were used. Wild-type and AMPK beta 1 knockout samples were subjected to SDS-PAGE. ab32112 and ab18058 (loading control to Vinculin) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-AMPK beta 1 antibody [Y367] (ab32112) at 1/5000 dilution

    Lane 1 : (A) : NIH 3T3.
    Lane 2 : (B) : Hela.
    Lane 3 : (C) : A431.
    Lane 4 : (D) : PC-12.


    Predicted band size : 30 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)
  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling AMPK beta 1 with purified ab32112 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

  • ab32112 at a 1:100 dilution staining AMPK beta 1 in human lung carcinoma, using Immunohistochemistry, Paraffin Embedded Tissue.
  • Overlay histogram showing HeLa cells stained with ab32112 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32112, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-AMPK beta 1 antibody [Y367] (ab32112)

This product has been referenced in:
  • Dzamko N  et al. AMPK beta1 deletion reduces appetite, preventing obesity and hepatic insulin resistance. J Biol Chem 285:115-22 (2010). Read more (PubMed: 19892703) »
  • Bendayan M  et al. Association of AMP-activated protein kinase subunits with glycogen particles as revealed in situ by immunoelectron microscopy. J Histochem Cytochem 57:963-71 (2009). Read more (PubMed: 19581628) »

See all 4 Publications for this product

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