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    ampk-gamma-1-antibody-y308-ab32508.pdf

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RecombinantRabMAb

Recombinant Anti-AMPK gamma 1 antibody [Y308] (ab32508)

  • Datasheet
  • SDS
Reviews (7) Submit a question References (22)

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Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
  • Flow Cytometry (Intracellular) - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
  • Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
  • Immunoprecipitation - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
  • Anti-AMPK gamma 1 antibody [Y308] (ab32508)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y308] to AMPK gamma 1
  • Suitable for: Flow Cyt (Intra), IP, WB
  • Reacts with: Human

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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: Flow Cyt (Intra), IP, WBmore details
    Unsuitable for: ICC or IHC
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, African green monkey
  • Immunogen

    Synthetic peptide within Human AMPK gamma 1 aa 300-400 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab218345)

  • Positive control

    • Jurkat whole cell lysate (ab7899). Flow Cyt (intra): HeLa cells
  • 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
    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.

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

    Y308
  • 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
    • Pathways and Processes
    • Metabolic signaling pathways
    • 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

    • Anti-AMPK gamma 1 antibody [Y308] - BSA and Azide free (ab239818)
  • Isotype control

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

    • Jurkat whole cell lysate (ab7899)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/100.

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

IP
Use at an assay dependent concentration.
WB (7)
1/1000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
Notes
Flow Cyt (Intra)
1/100.

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

IP
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 38 kDa).
Application notes
Is unsuitable for ICC or IHC.

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
    • Entrez Gene: 19082 Mouse
    • Entrez Gene: 25520 Rat
    • Omim: 602742 Human
    • SwissProt: P54619 Human
    • SwissProt: O54950 Mouse
    • SwissProt: P80385 Rat
    • Unigene: 530862 Human
    • Unigene: 6670 Mouse
    • Unigene: 11089 Rat
    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
    • 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 [Y308] (ab32508)
    Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)This image is courtesy of an anonymous Abreview
    All 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

    See Abreview

  • Flow Cytometry (Intracellular) - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
    Flow Cytometry (Intracellular) - Anti-AMPK gamma 1 antibody [Y308] (ab32508)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling AMPK gamma 1 with ab32508 at 1/100 dilution (1µg) (Red). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were fixed with4% paraformaldehyde. Rabbit monoclonal IgG (ab172730) was used as isotype control (Black). Unlabelled control: Cells without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Western blot - Anti-AMPK gamma 1 antibody [Y308] (ab32508)
    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)
    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.

  • Anti-AMPK gamma 1 antibody [Y308] (ab32508)
    Anti-AMPK gamma 1 antibody [Y308] (ab32508)

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

  • SDS download

  • Datasheet download

    Download

References (22)

Publishing research using ab32508? Please let us know so that we can cite the reference in this datasheet.

ab32508 has been referenced in 22 publications.

  • Olivier S  et al. Deletion of intestinal epithelial AMP-activated protein kinase alters distal colon permeability but not glucose homeostasis. Mol Metab 47:101183 (2021). PubMed: 33548500
  • Zhou F  et al. Jian-Pi-Yi-Shen Formula Ameliorates Oxidative Stress, Inflammation, and Apoptosis by Activating the Nrf2 Signaling in 5/6 Nephrectomized Rats. Front Pharmacol 12:630210 (2021). PubMed: 33841151
  • Yang SJ  et al. Regulation of AMPK Activity by CRBN Is Independent of the Thalidomide-CRL4CRBN Protein Degradation Axis. Pharmaceuticals (Basel) 14:N/A (2021). PubMed: 34073624
  • Maldonado M  et al. The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model. Aging (Albany NY) 13:16834-16858 (2021). PubMed: 34166224
  • 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). PubMed: 31241768
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-7 of 7 Abreviews or Q&A

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (Muscle fibers)
Gel Running Conditions
Reduced Denaturing (8% acrylamide)
Loading amount
50 µg
Specification
Muscle fibers
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Irene Tobias

Verified customer

Submitted Mar 21 2018

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% gradient gel)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Julie Kim

Verified customer

Submitted Apr 17 2017

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
African Green Monkey Cell lysate - whole cell (COS7)
Specification
COS7
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Apr 03 2014

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Odyssey block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Nov 25 2013

Western blot

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (10% tris glycine)
Sample
Mouse Cell lysate - other (mouse primary hepatocytes)
Specification
mouse primary hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted May 13 2013

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - nuclear (AMPK gama 1 infected cells)
Loading amount
20 µg
Specification
AMPK gama 1 infected cells
Gel Running Conditions
Reduced Denaturing (12% SDS PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Sep 07 2012

Western blot abreview for Anti-AMPK gamma 1 antibody [Y308]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Loading amount
20 µg
Specification
Skeletal muscle
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Read More

Mr. Jonas Treebak

Verified customer

Submitted Oct 30 2009

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