Recombinant Anti-AMPK gamma 1 antibody [Y307] (ab32382)
![Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)](https://www.abcam.com/ps/products/32/ab32382/Images/ab32382-332264-anti-ampk-gamma-1-antibody-y307-western-blot.jpg "Western blot - 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
Overview
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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.
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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
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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.
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
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
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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
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 |
|---|---|---|
| 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
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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
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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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 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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![Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)](https://www.abcam.com/ps/products/32/ab32382/Images/ab32382-44662-anti-ampk-gamma-1-antibody-y307-western-blot.jpg)
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)](https://www.abcam.com/ps/products/32/ab32382/Images/ab32382-44661-anti-ampk-gamma-1-antibody-y307-immunohistochemistry.jpg)
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.
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![Western blot - Anti-AMPK gamma 1 antibody [Y307] (ab32382)](https://www.abcam.com/ps/products/32/ab32382/Images/ab32382-44660-anti-ampk-gamma-1-antibody-y307-western-blot.jpg)
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
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![Anti-AMPK gamma 1 antibody [Y307] (ab32382)](https://www.abcam.com/ps/products/32/ab32382/Images/ab32382-5-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
References (2)
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
