Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

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Overview

  • Product name

    Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012]
    See all COX2 / Cyclooxygenase 2 primary antibodies

  • Description

    Rabbit monoclonal [EPR12012] to COX2 / Cyclooxygenase 2

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human COX2/ Cyclooxygenase 2 aa 550 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: P35354

  • Positive control

    • WB: A549 cell lysate and mouse spleen tissue lysate. IHC-P: Human colonic carcinoma, lung carcinoma, liver and colon tissues and rat kidney tissue lysate. IP: A549 cell lysate ICC/IF: U-87 MG cells

  • General notes

    A trial size is available to purchase for this antibody.

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab179800 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 - 1/5000. Predicted molecular weight: 69 kDa.
IP 1/10 - 1/100.
ICC/IF Use at an assay dependent concentration.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Mediates the formation of prostaglandins from arachidonate. May have a role as a major mediator of inflammation and/or a role for prostanoid signaling in activity-dependent plasticity.

    • Pathway

      Lipid metabolism; prostaglandin biosynthesis.

    • Sequence similarities

      Belongs to the prostaglandin G/H synthase family.
      Contains 1 EGF-like domain.

    • Post-translational
      modifications

      S-nitrosylation by NOS2 (iNOS) activates enzme activity. S-nitrosylation may take place on different Cys residues in addition to Cys-561.

    • Cellular localization

      Microsome membrane. Endoplasmic reticulum membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • COX 2 antibody
        • COX-2 antibody
        • COX2 antibody
        • Cyclooxygenase 2 antibody
        • Cyclooxygenase 2b antibody
        • Cyclooxygenase antibody
        • Cyclooxygenase-2 antibody
        • Cyclooxygenase2 antibody
        • EC 1.14.99.1 antibody
        • fj02a10 antibody
        • Glucocorticoid-regulated inflammatory cyclooxygenase antibody
        • Glucocorticoid-regulated inflammatory Prostaglandin G/H synthase antibody
        • GRIPGHS antibody
        • hCox 2 antibody
        • Macrophage activation-associated marker protein P71/73 antibody
        • OTTHUMP00000033524 antibody
        • PES-2 antibody
        • PGG/HS antibody
        • PGH synthase 2 antibody
        • PGH2_HUMAN antibody
        • PGHS 2 antibody
        • PGHS-2 antibody
        • PGHS2 antibody
        • PHS 2 antibody
        • PHS II antibody
        • PHS2 antibody
        • Prostaglandin endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) antibody
        • Prostaglandin endoperoxide synthase 2 antibody
        • Prostaglandin G/H synthase 2 antibody
        • Prostaglandin G/H synthase 2 precursor antibody
        • Prostaglandin G/H synthase and cyclooxygenase antibody
        • Prostaglandin G/H synthase antibody
        • Prostaglandin H2 synthase 2 antibody
        • prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) antibody
        • Prostaglandin-endoperoxide synthase 2 antibody
        • PTGS2 antibody
        • ptgs2a antibody
        • TIS10 antibody
        • TIS10 protein antibody
        • unp1239 antibody
        • wu:fj02a10 antibody
        see all

      Images

      • Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Lane 1: A549 cell lysate (20 µg)

        Lane 2: U-87 MG cell lysate (20 µg)

        Lane 3: HeLa wildtype cell lysate (20 µg)

        Lane 4: PTGS2 HeLa knockout cell lysate (20 µg)

        Lanes 1 - 4: Merged signal (red and green). Green - ab179800 observed at 75 kDa. Red - loading control, ab8245 observed at 37 kDa.

        ab179800 was shown to react with COX2 / Cyclooxygenase 2 in HeLa wildtype. Loss of signal was observed when knockout sample ab263795 was used. Wild-type and COX2 / Cyclooxygenase 2 knockout samples were subjected to SDS-PAGE. ab179800 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.
      • Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunocytochemistry/ Immunofluorescence analysis of U-87 MG (human glioblastoma-astrocytoma epithelial cell) cells labeling COX2 / Cyclooxygenase 2 with ab179800 at 1/50 dilution. ab150077 (AlexaFluor®488 Goat anti-Rabbit) at 1/1000 was used as secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as counterstain. Nuclie were stained blue with DAPI.

        Confocal image showing cytoplasmic staining in U-87 MG cell line.
        Negative control: MCF7 （PMID: 18199541）

      • Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution (Purified)

        Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate with 5% NFDM/TBST
        Lane 2 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate with 5% NFDM/TBST
        Lane 3 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate with 5% NFDM/TBST

        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: 69 kDa
        Observed band size: 72 kDa
        why is the actual band size different from the predicted?



        Exposure time

        Lane 1: 3.25 seconds
        Lane 2 and 3: 180 seconds

         

        The expression profile observed in HCT 116 and MCF7 are consistent with the literatures (PMID: 14739610, PMID: 24325753, PMID: 16997132).
        Negative control: HCT 116 (PMID: 14739610) and MCF7 (PMID: 24325753, PMID: 16997132)

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human colon tissue labeling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human liver tissue labelling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.

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

      • Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/5000 dilution (purified) + Mouse spleen tissue lysate at 20 µg

        Secondary
        HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/50000 dilution

        Predicted band size: 69 kDa
        Observed band size: 72 kDa why is the actual band size different from the predicted?



        Blocking and dilution buffer: 5% NFDM/TBST.

      • Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/5000 dilution (purified) + A549 whole cell lysate at 20 µg

        Secondary
        HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG at 1/50000 dilution

        Predicted band size: 69 kDa
        Observed band size: 72 kDa why is the actual band size different from the predicted?



        Blocking and dilution buffer: 5% NFDM/TBST.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      • Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        ab179800 (purified) at 1/30 immunoprecipitating COX2 in A549 whole cell lysate.

        Lane 1 (input): A549 whole cell lysate (10µg)

        Lane 2 (+): ab179800 + A549 whole cell lysate.

        Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab179800 in A549 whole cell lysate.

        For western blotting, HRP-conjugated anti-rabbit IgG, specific for the reduced form of IgG, was used as the secondary antibody (1/1500).

        Blocking buffer and concentration: 5% NFDM/TBST.

        Diluting buffer and concentration: 5% NFDM /TBST.

      • Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution (unpurified) + A549 cell lysate at 10 µg

        Predicted band size: 69 kDa

      • Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
        Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

        Western blot analysis on immunoprecipitation pellet from A549 cell lysate using unpurified ab179800.

      References

      This product has been referenced in:

      • Xu X  et al. Overexpression of lncRNA GAS5 suppresses prostatic epithelial cell proliferation by regulating COX-2 in chronic non-bacterial prostatitis. Cell Cycle 18:923-931 (2019). Read more (PubMed: 30892130) »
      • Liu Y  et al. Sinomenine retards LPS-elicited inflammation via down-regulating CCAT1 in HaCaT cells. Life Sci 233:116703 (2019). Read more (PubMed: 31356903) »
      See all 10 Publications for this product

      Publishing research using ab179800? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      Western blot abreview for Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Rat Tissue lysate - whole (Liver)
      Gel Running Conditions
      Reduced Denaturing (10% gel)
      Loading amount
      20 µg
      Specification
      Liver
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
      Read More

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      Verified customer

      Submitted Jun 12 2015

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