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  1. Link

    ampk-gamma-1-antibody-y307-ab32382.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-AMPK gamma 1 antibody [Y307] (ab32382)

  • Datasheet
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Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
  • Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
  • Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
  • Anti-AMPK gamma 1 antibody [Y307] (ab32382)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y307] to AMPK gamma 1
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)

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Overview

  • Product name

    Anti-AMPK gamma 1 antibody [Y307]
    See all AMPK gamma 1 primary antibodies
  • Description

    Rabbit monoclonal [Y307] to AMPK gamma 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human AMPK gamma 1 aa 1-100. The exact sequence is proprietary.

  • Epitope

    ab32382 reacts with an epitope located in the N terminal region of AMPK gamma 1.
  • Positive control

    • Jurkat cell lysate and HeLa cell lysate, human skeletal muscles
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y307
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Cardiovascular
    • Lipids / Lipoproteins
    • Fatty Acids
    • Synthesis
    • Signal Transduction
    • Signaling Pathway
    • Lipid Signaling
    • Other
    • Signal Transduction
    • Second Messenger
    • Nucleotide Messenger
    • cAMP
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
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    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Fatty acid oxidation

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-AMPK gamma 1 antibody [Y307] (ab205423)
    • Anti-AMPK gamma 1 antibody [Y307] - BSA and Azide free (ab247251)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)

Applications

Our Abpromise guarantee covers the use of ab32382 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 - 1/2000. Predicted molecular weight: 38 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    AMP/ATP-binding 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. Gamma non-catalytic subunit mediates binding to AMP, ADP and ATP, leading to activate or inhibit AMPK: AMP-binding results in allosteric activation of alpha catalytic subunit (PRKAA1 or PRKAA2) both by inducing phosphorylation and preventing dephosphorylation of catalytic subunits. ADP also stimulates phosphorylation, without stimulating already phosphorylated catalytic subunit. ATP promotes dephosphorylation of catalytic subunit, rendering the AMPK enzyme inactive.
  • Sequence similarities

    Belongs to the 5'-AMP-activated protein kinase gamma subunit family.
    Contains 4 CBS domains.
  • Domain

    The AMPK pseudosubstrate motif resembles the sequence around sites phosphorylated on target proteins of AMPK, except the presence of a non-phosphorylatable residue in place of Ser. In the absence of AMP this pseudosubstrate sequence may bind to the active site groove on the alpha subunit (PRKAA1 or PRKAA2), preventing phosphorylation by the upstream activating kinase STK11/LKB1.
    The CBS domains mediate binding to AMP, ADP and ATP. 2 sites bind either AMP or ATP, whereas a third site contains a tightly bound AMP that does not exchange. Under physiological conditions AMPK mainly exists in its inactive form in complex with ATP, which is much more abundant than AMP.
  • Post-translational
    modifications

    Phosphorylated by ULK1 and ULK2; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1, ULK2 and AMPK.
  • Target information above from: UniProt accession P54619 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5571 Human
    • Omim: 602742 Human
    • SwissProt: P54619 Human
    • Unigene: 530862 Human
    • Alternative names

      • 5' AMP activated protein kinase gamma 1 subunit antibody
      • 5' AMP activated protein kinase subunit gamma 1 antibody
      • 5''-AMP-activated protein kinase subunit gamma-1 antibody
      • AAKG1_HUMAN antibody
      • AMP activated protein kinase noncatalytic gamma 1 subunit antibody
      • AMPK gamma 1 chain antibody
      • AMPK gamma1 antibody
      • AMPK subunit gamma-1 antibody
      • AMPKg antibody
      • MGC8666 antibody
      • PRKAG 1 antibody
      • PRKAG1 antibody
      • Protein kinase AMP activated gamma 1 non catalytic subunit antibody
      • protein kinase, AMP-activated, noncatalytic gamma-1 antibody
      see all

    Images

    • Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      All lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : PRKAG1 (AMPK gamma 1) knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 38 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab32382 observed at 38 kDa. Red - loading control, Mouse anti Vinculin, observed at 125 kDa.

      ab32382 was shown to recognize AMPK gamma 1 in wild-type HAP1 cells as signal was lost at the expected MW in PRKAG1 (AMPK gamma 1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PRKAG1 (AMPK gamma 1) knockout samples were subjected to SDS-PAGE. Ab32382 and Mouse anti-Vinculin (ab9484 loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      All lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/2000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate

      Predicted band size: 38 kDa
      Observed band size: 37 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK gamma 1 antibody [Y307] (ab32382)

      This image shows paraffin embedded human skeletal muscle stained with 1/250 ab23282.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)This image is courtesy of an anonymous Abreview
      All lanes : Anti-AMPK gamma 1 antibody [Y307] (ab32382) at 1/2000 dilution

      Lane 1 : HeLa whole cell lystate - Control Si
      Lane 2 : HeLa whole cell lystate - treated with AMPK gamma 1 siRNA A for 36 hours
      Lane 3 : HeLa whole cell lystate - treated with AMPK gamma 1 siRNA B for 36 hours

      Secondary
      All lanes : HRP-conjugated Donkey polyclonal to rabbit IgG at 1/3000 dilution

      Predicted band size: 38 kDa

      See Abreview

    • Anti-AMPK gamma 1 antibody [Y307] (ab32382)
      Anti-AMPK gamma 1 antibody [Y307] (ab32382)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (2)

    Publishing research using ab32382? Please let us know so that we can cite the reference in this datasheet.

    ab32382 has been referenced in 2 publications.

    • Head SA  et al. Antifungal drug itraconazole targets VDAC1 to modulate the AMPK/mTOR signaling axis in endothelial cells. Proc Natl Acad Sci U S A 112:E7276-85 (2015). PubMed: 26655341
    • Preuss KD  et al. Inactivation of protein-phosphatase 2A causing hyperphosphorylation of autoantigenic paraprotein targets in MGUS/MM is due to an exchange of its regulatory subunits. Int J Cancer 135:2046-53 (2014). PubMed: 24676687

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-AMPK gamma 1 antibody [Y307]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    100000 cells
    Specification
    HeLa
    Treatment
    AMPK gamma1 siRNA for 36hrs
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 11 2010

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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