ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionNon-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 similaritiesBelongs to the 5'-AMP-activated protein kinase beta subunit family.
DomainThe 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.
5''-AMP-activated protein kinase subunit beta-1 antibody
5'-AMP-activated protein kinase beta-1 subunit antibody
AMP-activated protein kinase beta subunit antibody
AMP-ACTIVATED PROTEIN KINASE, NONCATALYTIC, BETA-1 antibody
AMP-activated, noncatalytic, beta-1 antibody
AMPK beta 1 chain antibody
AMPK subunit beta-1 antibody
Protein kinase AMP activated non catalytic subunit beta 1 antibody
protein kinase, AMP-activated, beta 1 non-catalytic subunit antibody
protein kinase, AMP-activated, noncatalytic, beta-1 antibody
Anti-AMPK beta 1 antibody [Y367] images
Western blot - Anti-AMPK beta 1 antibody [Y367] (ab32112)
Predicted band size : 30 kDa Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: AMPK beta 1 knockout HAP1 cell lysate Lane 3: HeLa cell lysate Lane 4: A431 cell lysate Lanes 1 - 4: Merged signal (red and green). Green - ab32112 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab32112 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. ab32112 and ab18058 (loading control to Vinculin) were both diluted 1/10000 and 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.
Western blot - AMPK beta 1 antibody [Y367] (ab32112)
All lanes : Anti-AMPK beta 1 antibody [Y367] (ab32112) at 1/5000 dilution
Lane 1 : (A) : NIH 3T3. Lane 2 : (B) : Hela. Lane 3 : (C) : A431. Lane 4 : (D) : PC-12.
Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling AMPK beta 1 with purified ab32112 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Overlay histogram showing HeLa cells stained with ab32112 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32112, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
References for Anti-AMPK beta 1 antibody [Y367] (ab32112)
This product has been referenced in:
Dzamko N et al. AMPK beta1 deletion reduces appetite, preventing obesity and hepatic insulin resistance. J Biol Chem285:115-22 (2010).
Read more (PubMed: 19892703) »
Bendayan M et al. Association of AMP-activated protein kinase subunits with glycogen particles as revealed in situ by immunoelectron microscopy. J Histochem Cytochem57:963-71 (2009).
Read more (PubMed: 19581628) »