

Anti-AMPK gamma 1 antibody [Y307] (ab32382)
- Datasheet
- References (1)
- Protocols
Overview
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Product nameAnti-AMPK gamma 1 antibody [Y307]
See all AMPK gamma 1 primary antibodies -
DescriptionRabbit monoclonal [Y307] to AMPK gamma 1
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC, Flow Cyt, IP, IHC-Pmore details
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Species reactivityReacts with: Human
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Immunogen
Synthetic peptide within Human AMPK gamma 1 aa 1-100. The exact sequence is proprietary.
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Epitopeab32382 reacts with an epitope located in the N terminal region of AMPK gamma 1.
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Positive control
- Jurkat cell lysate and HeLa cell lysate, human skeletal muscles
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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.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading...
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PurityProtein A purified
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ClonalityMonoclonal
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Clone numberY307
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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 |
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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. |
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IP | 1/50. | |
IHC-P | Use at an assay dependent concentration. |
Target
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FunctionAMP/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.
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Sequence similaritiesBelongs to the 5'-AMP-activated protein kinase gamma subunit family.
Contains 4 CBS domains. -
DomainThe 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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 5571 Human
- Omim: 602742 Human
- SwissProt: P54619 Human
- Unigene: 530862 Human
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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
see all
Images
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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 kDaLanes 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.
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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)This image shows paraffin embedded human skeletal muscle stained with 1/250 ab23282.
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Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)This image is courtesy of an anonymous AbreviewAll 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
Datasheets and documents
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) »