Overview

  • Product name

    Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free
    See all AMPK beta 1 primary antibodies
  • Description

    Rabbit monoclonal [Y367] to AMPK beta 1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IP, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human AMPK beta 1 aa 150-250. The exact sequence is proprietary.
    Database link: Q9Y478

  • General notes

    Ab239804 is the carrier-free version of ab32112. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab239804 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab239804 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
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 30 kDa.
IHC-P Use at an assay dependent concentration.

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

 

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

Images

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • 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 Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • Unpurified ab32112 at a 1:100 dilution staining AMPK beta 1 in human lung carcinoma, using Immunohistochemistry, Paraffin Embedded Tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

References

ab239804 has not yet been referenced specifically in any publications.

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