Overview

  • Product name
    Anti-AMPK beta 1 antibody [Y367]
    See all AMPK beta 1 primary antibodies
  • Description
    Rabbit monoclonal [Y367] to AMPK beta 1
  • Specificity
    This antibody is specific for human AMPK beta 1.
  • Tested applications
    Suitable 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

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 1/1000.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

Flow Cyt 1/800.

For unpurified use at 1/50. 

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

IP 1/40.

For unpurified use at 1/80. 

ICC/IF 1/500.

Target

  • Function
    Non-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 similarities
    Belongs to the 5'-AMP-activated protein kinase beta subunit family.
  • Domain
    The 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
    • 5'-AMP-activated protein kinase beta-1 subunit antibody
    • AAKB1_HUMAN antibody
    • AMP-activated protein kinase beta subunit 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
    • protein kinase, AMP-activated, beta 1 non-catalytic subunit antibody
    • protein kinase, AMP-activated, noncatalytic, beta-1 antibody
    see all

Anti-AMPK beta 1 antibody [Y367] images

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

    Lane 1 : Mouse brain lysates
    Lane 2 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 30 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling AMPK beta 1 with purified ab32112 at 1:800 dilution (1 ug/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - 0.1% Tween-20. Unlabeled control - Rabbit monoclonal IgG (Black).

  • ab32112 (purified) at 1:40 dilution (2ug) immunoprecipitating AMPK beta 1 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates.

    Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 10ug

    Lane 2 (+): ab32112 & NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32112 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

  • Anti-AMPK beta 1 antibody [Y367] (ab32112) at 1/20000 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 30 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST

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

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 30 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)

    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling AMPK beta 1 with purified ab32112 at 1:1000 dilution (0.85 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.



  • 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 - Unpurified ab32112 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.

    Unpurified 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 (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling AMPK beta 1 with purified ab32112 at 1:1000 dilution (0.85 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

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

    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.

  • Unpurified 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 unpurified 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:
  • Madiraju AK  et al. Argininosuccinate synthetase regulates hepatic AMPK linking protein catabolism and ureagenesis to hepatic lipid metabolism. Proc Natl Acad Sci U S A 113:E3423-30 (2016). Read more (PubMed: 27247419) »
  • 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) »

See all 5 Publications for this product

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