Name: Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012]
Brand: RabMAb
SKU: ab179800
Rating: 4 (1 reviews)

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Western blot - 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

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.
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 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, U-87 MG and HeLa cell lysates; mouse spleen 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
General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR12012
Isotype

IgG
Research areas

Alternative Versions
- Alexa Fluor® 647 Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab225273)
- Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (ab227528)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human PTGS2 (COX2 / Cyclooxygenase 2) knockout HeLa cell line (ab255420)
KO cell lysates
- Human PTGS2 (COX2 / Cyclooxygenase 2) knockout HeLa cell lysate (ab263795)
Positive Controls
Recombinant Protein
- Recombinant human COX2 / Cyclooxygenase 2 protein (ab198646)
Related Products
- Recombinant Human COX2 / Cyclooxygenase 2 protein (ab159279)

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 - 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.

Application notes

Is unsuitable for Flow Cyt.

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.
Target information above from: UniProt accession P35354 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 292547 Mouse
- Unigene: 44369 Rat
see all
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

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

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.

Predicted band size: 69 kDa

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 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.
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.
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) 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) 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) 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)

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)

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) 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) 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)

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)

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)

Western blot analysis on immunoprecipitation pellet from A549 cell lysate using unpurified ab179800.
Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (ab179800)

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet

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

ab179800 has been referenced in 20 publications.

Zhang J et al. Protocatechuic acid attenuates anterior cruciate ligament transection-induced osteoarthritis by suppressing osteoclastogenesis. Exp Ther Med 19:232-240 (2020). PubMed: 31853294
Jobbagy S et al. Nrf2 activation protects against lithium-induced nephrogenic diabetes insipidus. JCI Insight 5:N/A (2020). PubMed: 31941842
Deng QF et al. Cyclooxygenase-2 mediates gefitinib resistance in non-small cell lung cancer through the EGFR/PI3K/AKT axis. J Cancer 11:3667-3674 (2020). PubMed: 32284763
Shen Y et al. The roles of mechanosensitive ion channels and associated downstream MAPK signaling pathways in PDLC mechanotransduction. Mol Med Rep 21:2113-2122 (2020). PubMed: 32323761
Fang P et al. Quercetin Attenuates d-GaLN-Induced L02 Cell Damage by Suppressing Oxidative Stress and Mitochondrial Apoptosis via Inhibition of HMGB1. Front Pharmacol 11:608 (2020). PubMed: 32431618

View all Publications for this product

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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

Abcam user community

Verified customer

Submitted Jun 12 2015

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Related Products

Applications

Target

Function

Pathway

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (20)

Customer reviews and Q&As

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

Post-translational
modifications