Overview

  • Product nameAnti-Mark3 antibody [EPR633Y]
    See all Mark3 primary antibodies
  • Description
    Rabbit monoclonal [EPR633Y] to Mark3
  • Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Mark3 aa 600-700 (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: HeLa, K562 and NIH/3T3 cell lysate. ICC/IF: MCF-7 cells. Flow Cyt: HeLa cells. IP: K562 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

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

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab52626 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
WB 1/1000 - 1/2000. Detects a band of approximately 86 kDa.
IP 1/20 - 1/60.
Flow Cyt 1/30 - 1/1000.

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

ICC/IF 1/50 - 1/100.
  • Application notesIs unsuitable for IHC-P.
  • Target

    • FunctionInvolved in the specific phosphorylation of microtubule-associated proteins for tau, MAP2 and MAP4. Phosphorylates CDC25C on 'Ser-216'.
    • Tissue specificityUbiquitous.
    • Sequence similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. MARK subfamily.
      Contains 1 KA1 (kinase-associated) domain.
      Contains 1 protein kinase domain.
      Contains 1 UBA domain.
    • Information by UniProt
    • Database links
    • Alternative names
      • C-TAK1 antibody
      • Cdc25C associated protein kinase 1 antibody
      • Cdc25C-associated protein kinase 1 antibody
      • cTAK1 antibody
      • ELKL motif kinase 2 antibody
      • EMK-2 antibody
      • Emk2 antibody
      • ETK 1 antibody
      • KIAA4230 antibody
      • KP78 antibody
      • MAP microtubule affinity regulating kinase 3 antibody
      • MAP/microtubule affinity-regulating kinase 3 antibody
      • Mark3 antibody
      • MARK3_HUMAN antibody
      • Par 1a antibody
      • PAR1A antibody
      • Protein kinase STK10 antibody
      • Ser/Thr protein kinase PAR-1 antibody
      • Serine threonine protein kinase p78 antibody
      • Serine/threonine-protein kinase p78 antibody
      • SerThr protein kinase PAR 1 antibody
      see all

    Anti-Mark3 antibody [EPR633Y] images

    • 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 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.

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

      Observed band size : 86 kDa (why is the actual band size different from the predicted?)
      Blocking and dilution buffer: 5% NFDM/TBST
    • 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

      Observed band size : 86 kDa (why is the actual band size different from the predicted?)
      Blocking and dilution buffer: 5% NFDM/TBST
    • 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 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.

    • 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 as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • 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

      Observed band size : 86 kDa (why is the actual band size different from the predicted?)
    • 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.

    References for Anti-Mark3 antibody [EPR633Y] (ab52626)

    This product has been referenced in:
    • Fredriksson K  et al. Proteomic analysis of proteins surrounding occludin and claudin-4 reveals their proximity to signaling and trafficking networks. PLoS One 10:e0117074 (2015). Read more (PubMed: 25789658) »
    • Denison FC  et al. LKB1 is an essential regulator of spermatozoa release during spermiation in the mammalian testis. PLoS One 6:e28306 (2011). IP ; Mouse . Read more (PubMed: 22145035) »

    See all 2 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunoprecipitation
    Sample Mouse Cell lysate - whole cell (Primary cortical neurons)
    Total protein in input 50 µg
    Specification Primary cortical neurons
    Treatment Amyloid b peptide 20uM for 30 minutes
    Immuno-precipitation step Protein A
    Username

    Dr. Claire Thornton

    Verified customer

    Submitted Dec 10 2009

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