Recombinant Anti-AMPK gamma 1 antibody [Y308] (ab32508)
- Datasheet
- References (14)
- Protocols
Overview
-
Product name
Anti-AMPK gamma 1 antibody [Y308]
See all AMPK gamma 1 primary antibodies -
Description
Rabbit monoclonal [Y308] to AMPK gamma 1 -
Host species
Rabbit -
Specificity
ab32508 recognises 5’-AMP-activated protein kinase (AMPK). -
Tested applications
Suitable for: WB, Flow Cyt, IPmore details
Unsuitable for: ICC or IHC -
Species reactivity
Reacts with: Mouse, Rat, Human, African green monkey -
Immunogen
Synthetic peptide within Human AMPK gamma 1 aa 300-400 (C terminal). The exact sequence is proprietary.
(Peptide available asab218345) -
Positive control
- Jurkat whole cell lysate (ab7899).
-
General notes
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.
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, 0.05% BSA -
Concentration information loading... -
Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
Y308 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Isotype control
-
Positive Controls
Applications
Our Abpromise guarantee covers the use of ab32508 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 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa). | |
| Flow Cyt | 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| IP | 1/20 - 1/50. PubMed: 23612997 |
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
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
-
Database links
- Entrez Gene: 5571 Human
- Entrez Gene: 19082 Mouse
- Entrez Gene: 25520 Rat
- Omim: 602742 Human
- SwissProt: P54619 Human
- SwissProt: O54950 Mouse
- SwissProt: P80385 Rat
- Unigene: 530862 Human
see all -
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
-
![Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)](https://www.abcam.com/ps/products/32/ab32508/Images/ab32508-212149-anti-ampk-gamma-1-antibody-y308-western-blot.jpg)
Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)This image is courtesy of an anonymous AbreviewAll lanes : Anti-AMPK gamma 1 antibody [Y308] (ab32508) at 1/1000 dilution
All lanes : HEK293 whole cell lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Alexa Fluor® 690-conjugated Goat anti-rabbit IgG polyclonal at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 38 kDa
Exposure time: 5 minutes
-
![Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)](https://www.abcam.com/ps/products/32/ab32508/Images/ab32508-45061-anti-ampk-gamma-1-antibody-y308-western-blot.jpg)
Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)Anti-AMPK gamma 1 antibody [Y308] (ab32508) at 1/1000 dilution + Jurkat cell lysate
Predicted band size: 38 kDa
Observed band size: 38 kDa -
![Immunoprecipitation - Anti-AMPK gamma 1 antibody [Y308] (ab32508)](https://www.abcam.com/ps/products/32/ab32508/Images/ab32508-45060-anti-ampk-gamma-1-antibody-y308-immunoprecipitation.jpg)
Immunoprecipitation - Anti-AMPK gamma 1 antibody [Y308] (ab32508)AMPK gamma 1 was immunoprecipitated using 1 mg Jurkat whole cell extract, 0.2 ug of Rabbit monoclonal [Y308] to AMPK gamma 1and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Jurkat whole cell extractdiluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32508. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Bands: 37kDa: AMPK gamma 1.
References
This product has been referenced in:
- Dziewulska A et al. SCD1 regulates the AMPK/SIRT1 pathway and histone acetylation through changes in adenine nucleotide metabolism in skeletal muscle. J Cell Physiol 235:1129-1140 (2020). Read more (PubMed: 31241768) »
- Thomas EC et al. Isoform-specific AMPK association with TBC1D1 is reduced by a mutation associated with severe obesity. Biochem J 475:2969-2983 (2018). Read more (PubMed: 30135087) »
