Overview

  • Product name
    Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (HRP)
    See all AMPK alpha 1 + AMPK alpha 2 primary antibodies
  • Description
    Mouse monoclonal [34.2] to AMPK alpha 1 + AMPK alpha 2 (HRP)
  • Host species
    Mouse
  • Conjugation
    HRP
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Drosophila melanogaster
  • Immunogen

    Synthetic peptide corresponding to Fruit fly (Drosophila melanogaster) AMPK alpha 1 + AMPK alpha 2 (N terminal).
    Database link: Q13131

  • Positive control
    • WB: Rat heart and Human brain tissue lysates; HepG2 and HEK293 whole cell lysates. IHC-P: FFPE Human colon adenocarcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab202685 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
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 62,75,90 kDa (predicted molecular weight: 62 kDa).

Abcam recommends using 3% milk as the blocking agent.

Target

  • Function
    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. Regulates lipid synthesis by phosphorylating and inactivating lipid metabolic enzymes such as ACACA, ACACB, GYS1, HMGCR and LIPE; regulates fatty acid and cholesterol synthesis by phosphorylating acetyl-CoA carboxylase (ACACA and ACACB) and hormone-sensitive lipase (LIPE) enzymes, respectively. Regulates insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3. AMPK stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter SLC2A4/GLUT4 to the plasma membrane, possibly by mediating phosphorylation of TBC1D4/AS160. Regulates transcription and chromatin structure by phosphorylating transcription regulators involved in energy metabolism such as CRTC2/TORC2, FOXO3, histone H2B, HDAC5, MEF2C, MLXIPL/ChREBP, EP300, HNF4A, p53/TP53, SREBF1, SREBF2 and PPARGC1A. Acts as a key regulator of glucose homeostasis in liver by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. In response to stress, phosphorylates 'Ser-36' of histone H2B (H2BS36ph), leading to promote transcription. Acts as a key regulator of cell growth and proliferation by phosphorylating TSC2, RPTOR and ATG1: in response to nutrient limitation, negatively regulates the mTORC1 complex by phosphorylating RPTOR component of the mTORC1 complex and by phosphorylating and activating TSC2. In response to nutrient limitation, promotes autophagy by phosphorylating and activating ULK1. AMPK also acts as a regulator of circadian rhythm by mediating phosphorylation of CRY1, leading to destabilize it. May regulate the Wnt signaling pathway by phosphorylating CTNNB1, leading to stabilize it. Also has tau-protein kinase activity: in response to amyloid beta A4 protein (APP) exposure, activated by CAMKK2, leading to phosphorylation of MAPT/TAU; however the relevance of such data remains unclear in vivo. Also phosphorylates CFTR, EEF2K, KLC1, NOS3 and SLC12A1.
  • Sequence similarities
    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
    Contains 1 protein kinase domain.
  • Domain
    The AIS (autoinhibitory sequence) region some sequence similarity with the ubiquitin-associated domains and represses kinase activity.
  • Post-translational
    modifications
    Ubiquitinated.
    Phosphorylated at Thr-183 by STK11/LKB1 in complex with STE20-related adapter-alpha (STRADA) pseudo kinase and CAB39. Also phosphorylated at Thr-183 by CAMKK2; triggered by a rise in intracellular calcium ions, without detectable changes in the AMP/ATP ratio. CAMKK1 can also phosphorylate Thr-183, but at a much lower level. Dephosphorylated by protein phosphatase 2A and 2C (PP2A and PP2C). 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.
  • Cellular localization
    Cytoplasm. Nucleus. In response to stress, recruited by p53/TP53 to specific promoters.
  • Information by UniProt
  • Database links
  • Alternative names
    • 5''-AMP-activated protein kinase catalytic subunit alpha-1 antibody
    • AAPK1_HUMAN antibody
    • ACACA kinase antibody
    • Acetyl-CoA carboxylase kinase antibody
    • AMPK alpha 1 chain antibody
    • AMPK alpha 2 chain antibody
    • AMPK antibody
    • AMPK subunit alpha-1 antibody
    • AMPK1 antibody
    • AMPK2 antibody
    • HMGCR kinase antibody
    • Hydroxymethylglutaryl-CoA reductase kinase antibody
    • PRKAA antibody
    • PRKAA1 antibody
    • PRKAA2 antibody
    • Protein kinase AMP activated alpha 1 catalytic subunit antibody
    • Protein kinase AMP activated alpha 2 catalytic subunit antibody
    • Tau-protein kinase PRKAA1 antibody
    see all

Images

  • IHC image of AMPK alpha 1 + AMPK alpha 2 staining in a section of formalin-fixed paraffin-embedded human colon adenocarcinoma*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab202685, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (HRP) (ab202685) at 1/5000 dilution

    Lane 1 : Heart (Rat) Tissue Lysate
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 62 kDa
    Observed band size: 62,75,90 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab202685 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab202685 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab202685.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
For licensing inquiries, please contact partnerships@abcam.com

Sign up