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

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 as ab218345)
Positive control
- Jurkat cell lysate.
General notes

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.

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

Immunizing Peptide (Blocking)
- AMPK gamma 1 peptide (ab218345)
Positive Controls
- Jurkat whole cell lysate (ab7899)

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).Can be blocked with AMPK gamma 1 peptide (ab218345).
Flow Cyt		1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP		Use at an assay dependent concentration. PubMed: 23612997

Application notes

Is unsuitable for ICC or IHC.

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

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
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)

AMPK gamma 1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 10ug 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 70^oC; 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.

Datasheet

This product has been referenced in:

Bathgate KE et al. Muscle health and performance in monozygotic twins with 30 years of discordant exercise habits. Eur J Appl Physiol N/A:N/A (2018). Read more (PubMed: 30006671) »
Yavari A et al. Mammalian ?2 AMPK regulates intrinsic heart rate. Nat Commun 8:1258 (2017). Read more (PubMed: 29097735) »

See all 11 Publications for this product

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

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

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

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

Irene Tobias

Verified customer

Submitted Mar 21 2018

Abcam has not validated the combination of species/application used in this Abreview.

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

Julie Kim

Verified customer

Submitted Apr 17 2017

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

Abcam user community

Verified customer

Submitted Apr 03 2014

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

Abcam user community

Verified customer

Submitted Nov 25 2013

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

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

Abcam user community

Verified customer

Submitted May 13 2013

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

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

Abcam user community

Verified customer

Submitted Sep 07 2012

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

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

Mr. Jonas Treebak

Verified customer

Submitted Oct 30 2009

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

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

Overview

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Images

Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

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

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

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

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

Western blot

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

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