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RabMAb

Anti-AMPK alpha 1抗体[Y365] (ab32047)

概述

  • 产品名称
    Anti-AMPK alpha 1抗体[Y365]
    参阅全部 AMPK alpha 1 一抗
  • 描述
    兔单克隆抗体[Y365] to AMPK alpha 1
  • 特异性
    This antibody is specific for human AMPK alpha 1.
  • 经测试应用
    适用于: ICC/IF, Flow Cyt, WB, IHC-P, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, African green monkey
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human AMPK alpha 1 aa 500 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q13131

  • 阳性对照
    • WB: HeLa, HepG2, C6, NIH/3T3 and MCF-7 cell lysate. IHC-P: Human cervical carcinoma and lung carcinoma tissues. ICC/IF: MCF-7 cells. Flow Cyt: HeLa cells. IP: HeLa cell lysate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

应用

Our Abpromise guarantee covers the use of ab32047 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
ICC/IF 1/250.
Flow Cyt 1/100 - 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/5000. Predicted molecular weight: 63 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

IP 1/40 - 1/50.

靶标

  • 功能
    Responsible for the regulation of fatty acid synthesis by phosphorylation of acetyl-CoA carboxylase. It also regulates cholesterol synthesis via phosphorylation and inactivation of hormone-sensitive lipase and hydroxymethylglutaryl-CoA reductase. Appears to act as a metabolic stress-sensing protein kinase switching off biosynthetic pathways when cellular ATP levels are depleted and when 5'-AMP rises in response to fuel limitation and/or hypoxia. This is a catalytic subunit.
  • 序列相似性
    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. SNF1 subfamily.
    Contains 1 protein kinase domain.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 5 AMP activated protein kinase alpha 1catalytic subunit antibody
    • 5 AMP activated protein kinase catalytic alpha 1 chain antibody
    • 5' AMP activated protein kinase catalytic subunit alpha 1 antibody
    • 5'-AMP-activated protein kinase catalytic subunit alpha-1 antibody
    • AAPK1 antibody
    • AAPK1_HUMAN antibody
    • ACACA kinase antibody
    • acetyl CoA carboxylase kinase antibody
    • AI194361 antibody
    • AI450832 antibody
    • AL024255 antibody
    • AMP -activate kinase alpha 1 subunit antibody
    • AMP-activated protein kinase, catalytic, alpha -1 antibody
    • AMPK 1 antibody
    • AMPK alpha 1 antibody
    • AMPK alpha 1 chain antibody
    • AMPK antibody
    • AMPK subunit alpha-1 antibody
    • AMPK1 antibody
    • AMPKa1 antibody
    • AMPKalpha1 antibody
    • C130083N04Rik antibody
    • cb116 antibody
    • EC 2.7.11.1 antibody
    • HMG CoA reductase kinase antibody
    • HMGCR kinase antibody
    • hormone sensitive lipase kinase antibody
    • Hydroxymethylglutaryl CoA reductase kinase antibody
    • im:7154392 antibody
    • kinase AMPK alpha1 antibody
    • MGC33776 antibody
    • MGC57364 antibody
    • OTTHUMP00000161795 antibody
    • OTTHUMP00000161796 antibody
    • PRKAA 1 antibody
    • PRKAA1 antibody
    • Protein kinase AMP activated alpha 1 catalytic subunit antibody
    • SNF1-like protein AMPK antibody
    • SNF1A antibody
    • Tau protein kinase PRKAA1 antibody
    • wu:fa94c10 antibody
    see all

图片



  • Predicted band size : 63 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: AMPK alpha knockout HAP1 cell lysate (20 µg)
    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: HeLa knockout HAP1 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32047 observed at 63 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab32047 was shown to specifically react with AMPK alpha when AMPK alpha knockout samples were used. Wild-type and AMPK alpha knockout samples were subjected to SDS-PAGE. ab32047 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/2000 respectively 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling AMPK alpha 1 with purified ab32047 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of MCF7 cells labelling AMPK alpha 1 with purified ab32047 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry analysis of HeLa cells labelling AMPK alpha 1 with purified ab32047 at 1/150 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/2000 dilution (purified)

    Lane 1 : C6 cell lysate
    Lane 2 : NIH/3T3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 63 kDa
    Observed band size : 63 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/10000 dilution (purified)

    Lane 1 : MCF7 cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : K562 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 63 kDa
    Observed band size : 63 kDa

    Blocking and dilution buffer: 5% NFDM/TBST.

  • ab32047 (purified) at 1/40 immunoprecipitating AMPK alpha 1 in HeLa whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate (10µg)

    Lane 2 (+): ab32047 + HeLa whole cell lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32047 in HeLa whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/5000 dilution (unpurified)

    Lane 1 : MCF-7 cell lysate
    Lane 2 : HeLa cell lysate


    Predicted band size : 63 kDa
    Observed band size : 63 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analaysis of human cervical carcinoma tissue labelling AMPK alpha 1 with unpurified ab32047 at a dilution of 1/100.

  • Overlay histogram showing HeLa cells stained with unpurified ab32047 (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 (unpurified ab32047, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

文献

This product has been referenced in:
  • Ma D  et al. Cancer Cell Mitochondria Targeting by Pancratistatin Analogs is Dependent on Functional Complex II and III. Sci Rep 7:42957 (2017). WB . Read more (PubMed: 28220885) »
  • Han X  et al. AMPK activation protects cells from oxidative stress-induced senescence via autophagic flux restoration and intracellular NAD(+) elevation. Aging Cell 15:416-27 (2016). Read more (PubMed: 26890602) »

See all 29 Publications for this product

客户评价及客户问答

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain slide)
Specification
Brain slide
Username

Delphine Franssen

Verified customer

提交于 May 17 2017

Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% Gradient)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Julie Kim

Verified customer

提交于 May 05 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (human HepG2 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
human HepG2 cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Dec 05 2016

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

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