Anti-AMPK alpha 1 + AMPK alpha 2抗体[34.2] (ab80039)

概述

  • 产品名称Anti-AMPK alpha 1 + AMPK alpha 2抗体[34.2]
    参阅全部 AMPK alpha 1 + AMPK alpha 2 一抗
  • 描述
    小鼠单克隆抗体[34.2] to AMPK alpha 1 + AMPK alpha 2
  • 特异性Recognises both the alpha 1 and the alpha 2 AMPK
  • 经测试应用适用于: ELISA, WB, IP, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Drosophila melanogaster
  • 免疫原

    29 amino acid peptide designed within the N-terminal catalytic domains of Drosophila AMPK alpha 1 and 2

  • 阳性对照
    • This antibody gave a positive signal in the following lysates: rat heart tissue; human brain tissue; HepG2 whole cell; HEK293 whole cell.

应用

Our Abpromise guarantee covers the use of ab80039 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ELISA Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 62 kDa).

Abcam recommends using 3% milk as the blocking agent.

IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

靶标

  • 功能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.
  • 序列相似性Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
    Contains 1 protein kinase domain.
  • 结构域The AIS (autoinhibitory sequence) region some sequence similarity with the ubiquitin-associated domains and represses kinase activity.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm. Nucleus. In response to stress, recruited by p53/TP53 to specific promoters.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] 图像

  • All lanes : Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (ab80039) at 1 µg/ml

    Lane 1 : Heart (Rat) Tissue Lysate
    Lane 2 : Human brain tissue lysate - total protein (ab29466)
    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.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 62 kDa
    Observed band size : 62 kDa
    Additional bands at : 75 kDa,80 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
    Abcam recommends using milk as the blocking agent when running western blots of ab80039. This blot was produced using a 10% 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 ab80039 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
  • Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (ab80039) at 1/1000 dilution + Mouse 3T3-L1 adipocytes whole cell lysate at 25 µg

    Secondary
    HRP-conjugated goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 62 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an Abreview submitted by Brandon d'Eon

    See Abreview

  • All lanes : Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (ab80039) at 1/1000 dilution

    Lane 1 : Lysate of 0-4 hour Drosophila embryos expressing control shRNA.
    Lane 2 : Lysate of 0-4 hour Drosophila embryos expressing SNF1A shRNA.

    Secondary
    Sheep anti-mouse IgG conjugated to HRP at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 62 kDa


    Exposure time : 10 minutes

    Image courtesy of Richelle Sopko, Harvard University, U.S.A.

    See Abreview

Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (ab80039)参考文献

This product has been referenced in:
  • Liang D  et al. Therapeutic efficacy of apelin on transplanted mesenchymal stem cells in hindlimb ischemic mice via regulation of autophagy. Sci Rep 6:21914 (2016). WB ; Mouse . Read more (PubMed: 26902855) »
  • Omura J  et al. Protective Roles of Endothelial AMP-Activated Protein Kinase Against Hypoxia-Induced Pulmonary Hypertension in Mice. Circ Res 119:197-209 (2016). Read more (PubMed: 27217398) »

See all 9 Publications for this product

Product Wall

ab6006 is a suitable secondary antibody for both ab80039 and ab110304.

ab6006 detects the heavy and light chains of mouse IgG and IgM and IgA. ab80039 and ab110304 are both of the mouse IgG isotype.

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (13%)
Sample Human Cell lysate - whole cell (HCT116 human colon carcinoma cell line)
Specification HCT116 human colon carcinoma cell line
Treatment Control (Ctl) and 1 mM Hydroxyurea (HU) for 24 hrs
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

提交于 Aug 22 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (13%)
Sample Mouse Cell lysate - whole cell (MEF cell lysate)
Specification MEF cell lysate
Treatment Control (Ctl) and 1 mM Hydroxyurea (HU) for 24 hrs
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

提交于 Aug 22 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (10% gel, Tris-Glycine SDS Running Buffer)
Sample Mouse Cell lysate - whole cell (3T3-L1 adipocytes)
Specification 3T3-L1 adipocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Roger McLeod

Verified customer

提交于 Jun 23 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-12%)
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)
Specification 0-4hr embryos
Treatment control shRNA and SNF1A shRNA embryos
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Richelle Sopko

Verified customer

提交于 Dec 18 2013

I can confirm that ab6006 is suitable for use with antibodies ab119484, ab80039 and ab110304.

I can confirm that the two antibodies are very alike.
They both are mouse monoclonal antibodies that were produced by using the same immunogen.
But they are different clones and we do not know where exactly they bind in the 29 amino acid immu...

Read More
Application Western blot
Sample Mouse Tissue lysate - whole (heart)
Loading amount 40 µg
Specification heart
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Dec 20 2011

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