Anti-AMPK beta 1 antibody (ab79885)
Key features and details
- Rabbit polyclonal to AMPK beta 1
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-AMPK beta 1 antibody
See all AMPK beta 1 primary antibodies -
Description
Rabbit polyclonal to AMPK beta 1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human AMPK beta 1 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab91614) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; SHSY-5Y.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab79885 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications OrangutanApplication Abreviews Notes WB ICC/IF Notes Application notesICC/IF: Use at a concentration of 10 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 30 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.Target
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Function
Non-catalytic 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. Beta non-catalytic subunit acts as a scaffold on which the AMPK complex assembles, via its C-terminus that bridges alpha (PRKAA1 or PRKAA2) and gamma subunits (PRKAG1, PRKAG2 or PRKAG3). -
Sequence similarities
Belongs to the 5'-AMP-activated protein kinase beta subunit family. -
Domain
The glycogen-binding domain may target AMPK to glycogen so that other factors like glycogen-bound debranching enzyme or protein phosphatases can directly affect AMPK activity. -
Post-translational
modificationsPhosphorylated when associated with the catalytic subunit (PRKAA1 or PRKAA2). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK. - Information by UniProt
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Database links
- Entrez Gene: 5564 Human
- Entrez Gene: 100173201 Orangutan
- Omim: 602740 Human
- SwissProt: Q9Y478 Human
- SwissProt: Q5R801 Orangutan
- Unigene: 726001 Human
- Unigene: 741184 Human
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Alternative names
- 1300015D22Rik antibody
- 5''-AMP-activated protein kinase subunit beta-1 antibody
- 5'-AMP-activated protein kinase beta-1 subunit antibody
see all
Images
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Western blot - Anti-AMPK beta 1 antibody (ab79885)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: AMPK beta 1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab79885 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.ab79885 was shown to specifically react with AMPK beta 1 when AMPK beta 1 knockout samples were used. Wild-type and AMPK beta 1 knockout samples were subjected to SDS-PAGE. ab79885 at a concentration of 1ug/mL and ab18058 (loading control to Vinculin) at a dilution of 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-AMPK beta 1 antibody (ab79885)All lanes : Anti-AMPK beta 1 antibody (ab79885) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Additional bands at: 260 kDa, 46 kDa, 50 kDa, 72 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
The 35 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to human 5'-AMP-activated protein kinase subunit beta-1 (AMPK beta 1). -
Immunocytochemistry/ Immunofluorescence - Anti-AMPK beta 1 antibody (ab79885)ICC/IF image of ab79885 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79885, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2, and MCF-7 cells at 10µg/ml, and in 100% Methanol fixed (5 min) HeLa, HepG2, MCF-7 cells at 10µg/ml.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab79885 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-AMPK beta 1 antibody (ab79885)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: AMPK beta 1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab79885 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.ab79885 was shown to specifically react with AMPK beta 1 when AMPK beta 1 knockout samples were used. Wild-type and AMPK beta 1 knockout samples were subjected to SDS-PAGE. ab79885 at a concentration of 1ug/mL and ab18058 (loading control to Vinculin) at a dilution of 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-AMPK beta 1 antibody (ab79885)All lanes : Anti-AMPK beta 1 antibody (ab79885) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Additional bands at: 260 kDa, 46 kDa, 50 kDa, 72 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
The 35 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to human 5'-AMP-activated protein kinase subunit beta-1 (AMPK beta 1). -
Immunocytochemistry/ Immunofluorescence - Anti-AMPK beta 1 antibody (ab79885)ICC/IF image of ab79885 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79885, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2, and MCF-7 cells at 10µg/ml, and in 100% Methanol fixed (5 min) HeLa, HepG2, MCF-7 cells at 10µg/ml.
