Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) is a rabbit monoclonal antibody that is used to detect COX1 / Cyclooxygenase 1 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Specificity confirmed with COX1 / Cyclooxygenase 1 knockout cell line validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|
Human | Expected | Tested | Tested | Expected | Tested |
Mouse | Tested | Tested | Tested | Tested | Tested |
Rat | Expected | Expected | Expected | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/10 - 1/100 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 - 1/10000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/10 - 1/100 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/150 | Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 min. For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/150 | Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 min. For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/150 | Notes Heat up to 98 °C, below boiling, and then let cool for 10-20 min. For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Dual cyclooxygenase and peroxidase that plays an important role in the biosynthesis pathway of prostanoids, a class of C20 oxylipins mainly derived from arachidonate ((5Z,8Z,11Z,14Z)-eicosatetraenoate, AA, C20:4(n-6)), with a particular role in the inflammatory response. The cyclooxygenase activity oxygenates AA to the hydroperoxy endoperoxide prostaglandin G2 (PGG2), and the peroxidase activity reduces PGG2 to the hydroxy endoperoxide prostaglandin H2 (PGH2), the precursor of all 2-series prostaglandins and thromboxanes. This complex transformation is initiated by abstraction of hydrogen at carbon 13 (with S-stereochemistry), followed by insertion of molecular O2 to form the endoperoxide bridge between carbon 9 and 11 that defines prostaglandins. The insertion of a second molecule of O2 (bis-oxygenase activity) yields a hydroperoxy group in PGG2 that is then reduced to PGH2 by two electrons (PubMed:7947975). Involved in the constitutive production of prostanoids in particular in the stomach and platelets. In gastric epithelial cells, it is a key step in the generation of prostaglandins, such as prostaglandin E2 (PGE2), which plays an important role in cytoprotection. In platelets, it is involved in the generation of thromboxane A2 (TXA2), which promotes platelet activation and aggregation, vasoconstriction and proliferation of vascular smooth muscle cells (Probable). Can also use linoleate (LA, (9Z,12Z)-octadecadienoate, C18:2(n-6)) as substrate and produce hydroxyoctadecadienoates (HODEs) in a regio- and stereospecific manner, being (9R)-HODE ((9R)-hydroxy-(10E,12Z)-octadecadienoate) and (13S)-HODE ((13S)-hydroxy-(9Z,11E)-octadecadienoate) its major products (By similarity).
COX1, PTGS1, Prostaglandin G/H synthase 1, Cyclooxygenase-1, Prostaglandin H2 synthase 1, Prostaglandin-endoperoxide synthase 1, COX-1, PGH synthase 1, PGHS-1, PHS 1
Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) is a rabbit monoclonal antibody that is used to detect COX1 / Cyclooxygenase 1 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Specificity confirmed with COX1 / Cyclooxygenase 1 knockout cell line validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Cyclooxygenase 1 (COX-1) also known as COX1 cyclooxygenase-1 COX one and the COX1 protein is an important enzyme in the conversion of arachidonic acid to prostaglandins. It is a heme-containing dimer enzyme with a molecular weight of approximately 70 kDa. COX-1 is widely expressed in most tissues and cell types where it plays a significant role in maintaining homeostatic functions. This protein is constitutively active meaning it is often active under normal physiological conditions.
COX-1 is involved in producing prostaglandins that regulate a variety of normal physiological processes including gastric mucosal protection renal blood flow and platelet aggregation. Unlike COX-2 COX-1 is not induced by inflammatory stimuli and is not part of an inducible complex. It serves to maintain essential physiological functions in various organs and systems making its activity critical for cellular maintenance.
COX-1 is primarily involved in the prostaglandin biosynthesis pathway. It converts arachidonic acid into prostaglandin H2 a precursor for other prostaglandins and thromboxanes. Thromboxane A2 produced from this pathway plays an important role in platelet aggregation and vasoconstriction linking COX-1's functions with hemostatic processes. Another protein involved in this pathway is thromboxane synthase which further processes the products of COX-1 activity.
COX-1’s role connects it closely to conditions like peptic ulcers and cardiovascular diseases. Inhibition of COX-1 by nonsteroidal anti-inflammatory drugs (NSAIDs) can lead to gastric mucosal damage contributing to the development of peptic ulcers. Additionally due to its involvement in platelet aggregation COX-1 affects thrombotic diseases. COX-1's interactions with proteins such as COX-2 become relevant in inflammation and pain management where selective inhibition of COX-2 is sought to reduce adverse effects related to COX-1.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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ab109025 (purified) at 1:20 dilution (0.8μg) immunoprecipitating COX1 / Cyclooxygenase 1 in C2C12 whole cell lysate.
Lane 1 (input): C2C12 (Mouse myoblasts myoblast) whole cell lysate,10μg
Lane 2 (+): ab109025 & C2C12 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109025 in C2C12 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025)
Predicted band size: 68 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.
PBS instead of the primary antibody was used as the negative control.
All lanes: Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) at 1/10000 dilution
Lane 1: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: L6 (Rat skeletal muscle myoblast) whole cell lysates at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
All lanes: Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) at 1/2000 dilution
Lane 1: HaCaT (Human skin keratinocyte) whole cell lysates at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 68 kDa
Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling COX1 / Cyclooxygenase 1 with purified ab109025 at 1/100 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.
PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.
PBS instead of the primary antibody was used as the negative control.
Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma) labelling COX1 with purified ab109025 at 1/50. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only.
All lanes: Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) at 1/1000 dilution
Lane 1: NIH/3T3 cell lysate at 10 µg
Lane 2: HaCaT cell lysate at 10 µg
Lane 3: Neuro 2a cell lysate at 10 µg
Lane 4: C2C12 cell lysate at 10 µg
Lane 5: A431 cell lysate at 10 µg
Lane 6: L6 cell lysate at 10 µg
Predicted band size: 68 kDa
Unpurified ab109025 at 1/250 dilution staining COX1 / Cyclooxygenase 1 in human skin by immunohistochemistry, paraffin-embedded tissue.
Unpurified ab109025 at 1/100 dilution staining COX1 / Cyclooxygenase 1 in HeLa cells by Immunofluorescence.
Overlay histogram showing NIH/3T3 cells stained with unpurified ab109025 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109025, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
False colour image of Western blot: Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab109025 was shown to bind specifically to COX1 / Cyclooxygenase 1. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PTGS1 knockout cell line Human PTGS1 knockout A-431 cell line ab270477 (knockout cell lysate ab270500).
To generate this image, wild-type and PTGS1 knockout A431 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.
Lanes 1 - 4: Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] - BSA and Azide free (Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] - BSA and Azide free ab219375) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025) at 1/1000 dilution
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: Western blot - Human PTGS1 knockout A-431 cell lysate (ab270500)
Lane 2: PTGS1 knockout A431 cell lysate at 20 µg
Lane 2: Western blot - Human PTGS1 knockout A-431 cell line (Human PTGS1 knockout A-431 cell line ab270477)
Lane 3: C2C12 cell lysate at 20 µg
Lane 4: MOLT-4 cell lysate at 20 µg
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 70 kDa
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