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.
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 : RAW 264.7 Control LPS (0 ng/mL, 4 h) cell lysate
Lane 2 : RAW 264.7 Treated LPS (100 ng/mL, 4 h) cell lysate
Lane 3 : Wild-type A549 ab277305 cell lysate
Lane 4 : PTGS2 knockout A549 ab283802 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 69 kDaWestern blot: Anti-PTGS2 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 PTGS2. A band was observed at 69 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PTGS2 knockout cell line. To generate this image, wild-type and PTGS2 knockout RAW 264.7 cell lysates were analysed. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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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) 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 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) 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) 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) 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) 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) 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) 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
- Gong M et al. PHLDA1 knockdown inhibits inflammation and oxidative stress by regulating JNK/ERK pathway, and plays a protective role in sepsis-induced acute kidney injury. Allergol Immunopathol (Madr) 50:1-9 (2022). PubMed: 36335439
- 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