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    mark3-antibody-epr633y-ab52626.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microtubules MT Associated Proteins MARK
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Mark3 antibody [EPR633Y] (ab52626)

  • Datasheet
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Immunoprecipitation - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Immunocytochemistry/ Immunofluorescence - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Flow Cytometry (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Flow Cytometry (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)
  • Anti-Mark3 antibody [EPR633Y] (ab52626)

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
  • Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Anti-AK2 antibody [EPR11388(B)] (ab166901)
Protein
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Recombinant human Mark3 protein (ab60012)

View more associated products

Overview

  • Product name

    Anti-Mark3 antibody [EPR633Y]
    See all Mark3 primary antibodies
  • Description

    Rabbit monoclonal [EPR633Y] to Mark3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt (Intra), ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide 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 (intra): HeLa cells. IP: K562 cells.
  • General notes

    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.

    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 this species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR633Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • MT Associated Proteins
    • MARK
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases

Associated products

  • Alternative Versions

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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • NIH/3T3 whole cell lysate (ab7179)
    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant human Mark3 protein (ab60012)

Applications

The Abpromise guarantee

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)
1/20 - 1/60.
Flow Cyt (Intra)
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.
Notes
WB
1/1000 - 1/2000. Detects a band of approximately 86 kDa.
IP
1/20 - 1/60.
Flow Cyt (Intra)
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 notes
Is unsuitable for IHC-P.

Target

  • Function

    Involved in the specific phosphorylation of microtubule-associated proteins for tau, MAP2 and MAP4. Phosphorylates CDC25C on 'Ser-216'.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Belongs 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.
  • Target information above from: UniProt accession P27448 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4140 Human
    • Entrez Gene: 17169 Mouse
    • Omim: 602678 Human
    • SwissProt: P27448 Human
    • SwissProt: Q03141 Mouse
    • Unigene: 35828 Human
    • Unigene: 260504 Mouse
    • Unigene: 408955 Mouse
    • Unigene: 420865 Mouse
    see all
  • 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

Images

  • Immunoprecipitation - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Immunoprecipitation - Anti-Mark3 antibody [EPR633Y] (ab52626)

    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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)

    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 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] (ab52626)
    Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
    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

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



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
    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 - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Immunocytochemistry/ Immunofluorescence - Anti-Mark3 antibody [EPR633Y] (ab52626)

    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 (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Flow Cytometry (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)

    Intracellular Flow Cytometry analysis ofHeLa cells labelling Mark3 with purified ab52626 at a dilution of 1/50 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluorr® 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] (ab52626)
    Western blot - Anti-Mark3 antibody [EPR633Y] (ab52626)
    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?

  • Flow Cytometry (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)
    Flow Cytometry (Intracellular) - Anti-Mark3 antibody [EPR633Y] (ab52626)

    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 Fluorr® 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] (ab52626)
    Anti-Mark3 antibody [EPR633Y] (ab52626)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (2)

Publishing research using ab52626? Please let us know so that we can cite the reference in this datasheet.

ab52626 has been referenced in 2 publications.

  • 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). 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 . PubMed: 22145035

Customer reviews and Q&As

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Immunoprecipitation abreview for Anti-Mark3 antibody [EPR633Y]

Excellent
Abreviews
Abreviews
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Claire Thornton

Verified customer

Submitted Dec 10 2009

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