Recombinant Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12012] to COX2 / Cyclooxygenase 2
- Suitable for: WB, IP, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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 -
Specificity
Stimulation is required to allow detection of the COX2 protein in some cell lines and tissues. It is better to use a positive control side by side when testing.
Rat species is recommended based on IHC result, we do not guarantee WB, IP and ICC/IF for Rat. -
Tested applications
Suitable for: WB, IP, ICC/IF, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, U-87 MG and HeLa cell lysates; mouse spleen tissue lysate, PTGS2 (COX2 / Cyclooxygenase 2) KO A549 (Human lung carcinoma epithelial cell) cell lysate, Wild-type A549 cell lysate, U-87 MG whole cell lysate, MCF7 whole cell lysate. Mouse B16-F10 and Raw 264.7 whole cell lysate. Mouse retina, hippocampus, heart and kidney tissue lysate. IHC-P: Human colonic carcinoma, lung carcinoma, liver and colon tissues: rat kidney tissue; mouse kidney and liver tissue. IP: A549 cell lysate ICC: U-87 MG cells
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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
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12012 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB | (1) |
1/1000 - 1/5000. Predicted molecular weight: 69 kDa.
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IP |
1/10 - 1/100.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (2) |
1/100 - 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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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 - 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsS-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
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Database links
- Entrez Gene: 5743 Human
- Entrez Gene: 19225 Mouse
- Entrez Gene: 29527 Rat
- Omim: 600262 Human
- SwissProt: P35354 Human
- SwissProt: Q05769 Mouse
- SwissProt: P35355 Rat
- Unigene: 196384 Human
see all -
Alternative names
- COX 2 antibody
- COX-2 antibody
- COX2 antibody
see all
Images
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : B16-F10 (Mouse skin melanoma) whole cell lysate
Lane 2 : Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Mouse retina tissue lysate
Lane 4 : Mouse hippocampus tissue lysate
Lane 5 : Mouse heart tissue lysate
Lane 6 : Mouse kidney tissue lysate
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: 60 secondsBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
COX2 is expressed at a low level in Raw264.7, mouse retina, hippocampus, heart, kidney etc. (PMID: 22015457, PMID: 26001832, PMID: 23045674, PMID: 33737575).
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : PTGS2 (COX2 / Cyclooxygenase 2) KO A549 (Human lung carcinoma epithelial cell) cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lane 2 : Wild-type A549 cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/100000 dilution
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 74 kDa why is the actual band size different from the predicted?Negative control: MCF7 (PMID: 24325753, PMID: 16997132)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
False colour image of Western blot: Anti- COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab179800 was shown to bind specifically to COX2 / Cyclooxygenase 2. Target band was observed at 74 kDa in wild-type A549 cell lysates with no signal observed at this size in COX2 / Cyclooxygenase 2 knockout cell line ab280802. To generate this image, wild-type and COX2 / Cyclooxygenase 2 knockout A549 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : PTGS2 knockout A549 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDaFalse colour image of Western blot: Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab179800 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 75 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). To generate this image, wild-type and PTGS2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution
Lane 1 : A549 cell lysate
Lane 2 : U-87 MG cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : PTGS2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDaLanes 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 wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255420 (knockout cell lysate 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain. -
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) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: secondary antibody only control. Hematoxylin was used as a counterstain. -
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 secondsThe 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) 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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800) at 1/1000 dilution (unpurified) + A549 cell lysate at 10 µg
Predicted band size: 69 kDa -
Western blot analysis on immunoprecipitation pellet from A549 cell lysate using unpurified ab179800.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (87)
ab179800 has been referenced in 87 publications.
- Zhou WZ et al. LncRNA-CASC15 knockdown inhibits the progression of esophageal squamous cell carcinoma through targeting miR-33a-5p/PTGS2 axis. Histol Histopathol 38:223-232 (2023). PubMed: 36111503
- Sun L et al. Repetitive Transcranial Magnetic Stimulation Reduces Neuronal Loss and Neuroinflammation in Parkinson?s Disease Through the miR-195a-5p/CREB Axis. Turk Neurosurg 33:229-237 (2023). PubMed: 36300577
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- Li L et al. Effect of Azelaic Acid on Psoriasis Progression Investigated Based on Phosphatidylinositol 3-Kinase (PI3K)/Protein Kinase B (AKT) Signaling Pathway. Clin Cosmet Investig Dermatol 15:2523-2534 (2022). PubMed: 36447569
- Chu Y et al. TRPC5 mediates endothelium-dependent contraction in the carotid artery of diet-induced obese mice. Hypertens Res 45:1945-1953 (2022). PubMed: 36123395