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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-COMT antibody [EPR6490] (ab126618)

  • Datasheet
  • SDS
Reviews (1) Submit a question References (5)

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Western blot - Anti-COMT antibody [EPR6490] (ab126618)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)
  • Western blot - Anti-COMT antibody [EPR6490] (ab126618)
  • Western blot - Anti-COMT antibody [EPR6490] (ab126618)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)
  • Anti-COMT antibody [EPR6490] (ab126618)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6490] to COMT
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

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Knockout
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Human COMT knockout HEK-293T cell lysate (ab257396)

View more associated products

Overview

  • Product name

    Anti-COMT antibody [EPR6490]
    See all COMT primary antibodies
  • Description

    Rabbit monoclonal [EPR6490] to COMT
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide corresponding to Human COMT. Human COMT is a soluble isoform.
    Database link: P21964

  • Positive control

    • WB: HEK-293T, HAP1, A431, MOLT4 and MCF7 cell lysates; Human fetal liver lysate. IHC-P: Human liver and kidney tissues.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR6490
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Other
    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-COMT antibody [EPR6490] - BSA and Azide free (ab248128)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human COMT knockout HEK-293T cell line (ab266537)
  • KO cell lysates

    • Human COMT knockout HEK-293T cell lysate (ab257396)
  • Positive Controls

    • A-431 membrane extract lysate (ab30134)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab126618 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).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 24-30 kDa (predicted molecular weight: 30 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF 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.
    • Target information above from: UniProt accession P21964 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 1312 Human
      • Entrez Gene: 12846 Mouse
      • Entrez Gene: 24267 Rat
      • Omim: 116790 Human
      • SwissProt: P21964 Human
      • SwissProt: O88587 Mouse
      • SwissProt: P22734 Rat
      • Unigene: 370408 Human
      • Unigene: 704514 Human
      • Unigene: 100940 Mouse
      • Unigene: 220 Rat
      see all
    • 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 [EPR6490] (ab126618)
      Western blot - Anti-COMT antibody [EPR6490] (ab126618)
      All lanes : Anti-COMT antibody [EPR6490] (ab126618) 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: 24-28 kDa why is the actual band size different from the predicted?



      Lanes 1-2: Merged signal (red and green). Green - ab126618 observed at 24-28 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab126618 Anti-COMT antibody [EPR6490] 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. ab126618 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.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)

      ab126618, at 1/50 dilution, staining COMT in Formalin-fixed, Paraffin-embedded Human liver tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-COMT antibody [EPR6490] (ab126618)
      Western blot - Anti-COMT antibody [EPR6490] (ab126618)
      All lanes : Anti-COMT antibody [EPR6490] (ab126618) at 1/1000 dilution

      Lane 1 : Wild type HAP1 whole cell lysate at 40 µg
      Lane 2 : COMT knockout HAP1 whole cell lysate at 40 µg
      Lane 3 : A431 whole cell lysate at 20 µg
      Lane 4 : MCF7 whole cell lysate at 20 µg

      Predicted band size: 30 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab126618 observed at 24/28 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab126618 was shown to specifically react with COMT when COMT knockout samples were used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. Samples were incubated with ab126618 and ab8245 (Mouse anti-GAPDH loading control) overnight at 4°C at 1/1000 and 1/10000 dilutions respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-COMT antibody [EPR6490] (ab126618)
      Western blot - Anti-COMT antibody [EPR6490] (ab126618)
      All lanes : Anti-COMT antibody [EPR6490] (ab126618) at 1/1000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : A431 cell lysate
      Lane 3 : MOLT4 cell lysate
      Lane 4 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 30 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COMT antibody [EPR6490] (ab126618)

      ab126618, at 1/50 dilution, staining COMT in Formalin-fixed, Paraffin-embedded Human kidney tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Anti-COMT antibody [EPR6490] (ab126618)
      Anti-COMT antibody [EPR6490] (ab126618)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab126618? Please let us know so that we can cite the reference in this datasheet.

    ab126618 has been referenced in 5 publications.

    • Winner BM  et al. Metabolism of Dopamine in Nucleus Accumbens Astrocytes Is Preserved in Aged Mice Exposed to MPTP. Front Aging Neurosci 9:410 (2017). PubMed: 29311899
    • Cooper TK  et al. Coati Bodies: Cytoplasmic Hyaline Globules in the Ganglionic Neurons of Aging Captive Coatis. Vet Pathol 54:851-854 (2017). PubMed: 28578627
    • Doan KV  et al. FoxO1 in dopaminergic neurons regulates energy homeostasis and targets tyrosine hydroxylase. Nat Commun 7:12733 (2016). PubMed: 27681312
    • Stefanovic B  et al. Melatonin mediated antidepressant-like effect in the hippocampus of chronic stress-induced depression rats: Regulating vesicular monoamine transporter 2 and monoamine oxidase A levels. Eur Neuropsychopharmacol 26:1629-37 (2016). WB ; Rat . PubMed: 27499503
    • Wakuda T  et al. Perinatal asphyxia alters neuregulin-1 and COMT gene expression in the medial prefrontal cortex in rats. Prog Neuropsychopharmacol Biol Psychiatry 56:149-54 (2015). PubMed: 25194460

    Customer reviews and Q&As

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    Immunohistochemistry (Frozen sections) abreview for Anti-COMT antibody [EPR6490]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Liver)
    Permeabilization
    No
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 25 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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