Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-COMT antibody [EPR6491(B)] (ab124813)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6491(B)] to COMT
  • Suitable for: WB, Flow Cyt
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-COMT antibody [EPR6491(B)]
    See all COMT primary antibodies

  • Description

    Rabbit monoclonal [EPR6491(B)] to COMT

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cytmore details
    Unsuitable for: ICC,IHC-P or IP

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Synthetic peptide within Human COMT aa 250-350. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293T, HAP1, A431 and MOLT4 cell lysates; Human fetal liver lysate. Flow Cyt: A431 cells.

  • General notes

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

     

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab124813 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 - 1/10000. Detects a band of approximately 24-30 kDa (predicted molecular weight: 30 kDa).
Flow Cyt 1/10 - 1/100.

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

 
  • Application notes
    Is unsuitable for ICC,IHC-P or IP.

    • Target

    • Function

      Catalyzes the O-methylation, and thereby the inactivation, of catecholamine neurotransmitters and catechol hormones. Also shortens the biological half-lives of certain neuroactive drugs, like L-DOPA, alpha-methyl DOPA and isoproterenol.

    • Tissue specificity

      Brain, liver, placenta, lymphocytes and erythrocytes.

    • Sequence similarities

      Belongs to the mammalian catechol-O-methyltransferase family.

    • Post-translational
      modifications

      The N-terminus is blocked.

    • Cellular localization

      Cytoplasm and Cell membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • Catechol O methyltransferase antibody
        • Catechol O-methyltransferase antibody
        • COMT antibody
        • COMT_HUMAN antibody
        • EC 2.1.1.6 antibody
        see all

      Images

      • Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        All lanes : Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

        Lane 1 : Wild-type HEK-293T cell lysate
        Lane 2 : COMT knockout HEK-293T cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 30 kDa
        Observed band size: 28 kDa
        why is the actual band size different from the predicted?



        Lanes 1-2: Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control ab8245 observed at 37 kDa.

         ab124813 Anti-COMT antibody [EPR6491(B)] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab124813 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

         

      • Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        All lanes : Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

        Lane 1 : Wild-type HAP1 whole cell lysate
        Lane 2 : COMT knockout HAP1 whole cell lysate
        Lane 3 : A431 whole cell lysate
        Lane 4 : MOLT4 whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Predicted band size: 30 kDa



        Lanes 1 - 4: Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control, ab18058, observed at 130 kDa.

        ab124813 was shown to recognize COMT when COMT knockout samples were used, along with additional cross-reactive bands. Wild-type and COMT knockout samples were subjected to SDS-PAGE.  Ab124813 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813)
        All lanes : Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

        Lane 1 : Human fetal liver lysate
        Lane 2 : A431 cell lysate
        Lane 3 : MOLT4 cell lysate

        Lysates/proteins at 10 µg per lane.

        Developed using the ECL technique.

        Predicted band size: 30 kDa

      • Flow Cytometry - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Flow Cytometry - Anti-COMT antibody [EPR6491(B)] (ab124813)
        ab124813, at 1/10 dilution staining COMT in A431 cells, (Red) compared to a nonspecific negative control antibody (Green) by Flow Cytometry.
      • Other - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Other - Anti-COMT antibody [EPR6491(B)] (ab124813)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD

      References (0)

      Publishing research using ab124813? Please let us know so that we can cite the reference in this datasheet.

      ab124813 has not yet been referenced specifically in any publications.

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