Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y365] to AMPK alpha 1 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free
    See all AMPK alpha 1 primary antibodies

  • Description

    Rabbit monoclonal [Y365] to AMPK alpha 1 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody is specific for human AMPK alpha 1. This antibody shows low affinity on mouse and rat samples.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HepG2, C6, NIH/3T3 and MCF-7 cell lysate. Mouse liver, brain, retina, and skeletal muscle tissue lysates. IHC-P: Human cervical carcinoma and lung carcinoma tissues. ICC/IF: MCF-7 cells. Flow Cyt: HeLa cells. IP: HeLa cell lysate.

  • General notes

    ab210714 is the carrier-free version of ab32047. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab210714 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Immunocytochemistry/ Immunofluorescence - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Immunocytochemistry/ Immunofluorescence - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Flow Cytometry - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Flow Cytometry - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Immunoprecipitation - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Immunoprecipitation - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Western blot - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Western blot - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/500 dilution

    Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 3 : Mouse liver tissue lysate
    Lane 4 : Mouse brain tissue lysate
    Lane 5 : Mouse kidney tissue lysate
    Lane 6 : Mouse retina tissue lysate
    Lane 7 : Mouse skeletal muscle tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 63 kDa
    Observed band size: 63 kDa
    Additional bands at: 40 kDa (possible non-specific binding)


    Exposure time: 3 minutes


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    This antibody shows low affinity on mouse samples. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Western blot - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Western blot - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    All lanes : Anti-AMPK alpha 1 antibody [Y365] (ab32047) at 1/20000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 5 : C6 (Rat glial tumor glial cell) whole cell lysate
    Lane 6 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
    Lane 7 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

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


    Exposure time: 180 seconds


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    This antibody shows low affinity on mouse and rat samples.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Flow Cytometry - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Flow Cytometry - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32047).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)
    Anti-AMPK alpha 1 antibody [Y365] - BSA and Azide free (ab210714)

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