Overview

  • Product name
  • Description
    Rabbit polyclonal to AMPK beta 1
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human AMPK beta 1.

    (Peptide available as ab91614.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; SHSY-5Y.

Properties

Applications

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.

Application Abreviews Notes
WB
ICC/IF
  • Application notes
    ICC/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

    • 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
      modifications
      Phosphorylated 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
    • Database links
    • Alternative names
      • 1300015D22Rik antibody
      • 5''-AMP-activated protein kinase subunit beta-1 antibody
      • 5'-AMP-activated protein kinase beta-1 subunit antibody
      • AAKB1_HUMAN antibody
      • AMP-activated protein kinase beta subunit antibody
      • AMP-ACTIVATED PROTEIN KINASE, NONCATALYTIC, BETA-1 antibody
      • AMP-activated, noncatalytic, beta-1 antibody
      • AMPK antibody
      • AMPK beta 1 chain antibody
      • AMPK subunit beta-1 antibody
      • AMPK-BETA-1 antibody
      • AMPKb antibody
      • AU021155 antibody
      • E430008F22 antibody
      • HAMPKb antibody
      • MGC17785 antibody
      • PRKAB1 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
      see all

    Images



    • 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 (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.

    • 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
      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 minutesThe 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).
    • 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.

    References

    ab79885 has not yet been referenced specifically in any publications.

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