Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR633Y] to Mark3 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), ICC, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Mark3 antibody [EPR633Y] - BSA and Azide free
    See all Mark3 primary antibodies

  • Description

    Rabbit monoclonal [EPR633Y] to Mark3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC, WB, IPmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • General notes

    ab247326 is the carrier-free version of ab52626.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

Images

  • Immunoprecipitation - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Immunoprecipitation - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)

    ab52626&

    44; the same antibody clone in a different buffer formulation.

    >ab52626 (purified) at a dilution of 1/20 immunoprecipitating Mark3 in K562 whole cell lysate.

    Lane 1 (input): K562 whole cell lysate (10µg).

    Lane 2 (+): ab52626 + K562 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52626 in K562 whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/1000). Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)

    This data was developed using ab52626, the same antibody clone in a different buffer formulation.

    Lane 1: Wild-type HAP1 cell lysate (40 µg)

    Lane 2: MARK3 knockout HAP1 cell lysate (40 µg)

    Lane 3: HeLa cell lysate (20 µg)

    Lane 4: K562 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab52626 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab52626 was shown to recognize Mark3 when Mark3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Mark3 knockout samples were subjected to SDS-PAGE. ab52626 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1:10,000 dilution 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 hour at room temperature before imaging.

  • Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    All lanes : Anti-Mark3 antibody [EPR633Y] (ab52626) at 1/2000 dilution (purified)

    Lane 1 : K562 whole cell lysate
    Lane 2 : HeLa whole cell 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: 87 kDa
    Observed band size: 86 kDa why is the actual band size different from the predicted?



    This data was developed using ab52626, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Anti-Mark3 antibody [EPR633Y] (ab52626) at 10 µg (purified) + NIH/3T3 whole cell lysate at 10 µg

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

    Predicted band size: 87 kDa
    Observed band size: 86 kDa why is the actual band size different from the predicted?



    This data was developed using ab52626, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Immunocytochemistry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    This data was developed using ab52626, the same antibody clone in a different buffer formulation.Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Mark3 with purified ab52626 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) 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 (1/1000) were also used. Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000). Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
  • Flow Cytometry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Flow Cytometry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    This data was developed using ab52626, the same antibody clone in a different buffer formulation.Flow Cytometry analysis of HeLa cells labelling Mark3 with purified ab52626 at a dilution of 1/50 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
  • Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Western blot - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Anti-Mark3 antibody [EPR633Y] (ab52626) at 1/2000 dilution (unpurified) + HeLa cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 87 kDa
    Observed band size: 86 kDa why is the actual band size different from the predicted?



    This data was developed using ab52626, the same antibody clone in a different buffer formulation.

  • Flow Cytometry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Flow Cytometry - Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    This data was developed using ab52626, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with unpurified ab52626 (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 ab52626, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 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.
  • Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)
    Anti-Mark3 antibody [EPR633Y] - BSA and Azide free (ab247326)

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