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, ICC, Flow Cyt, IP, 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.

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

    This product is a recombinant rabbit monoclonal antibody.

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.
ICC 1/50 - 1/100.
Flow Cyt 1/100.

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

IP 1/50.
IHC-P Use at an assay dependent concentration.

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.
  • Information by UniProt
  • Database links
  • 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

  • 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.

  • 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?

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

References

This product has been referenced in:
  • 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). Read more (PubMed: 26655341) »
See 1 Publication for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
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

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Verified customer

Submitted Feb 11 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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