AB325256

Human COX2/Cyclooxygenase 2 ELISA Kit

SimpleStep
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Human COX2/Cyclooxygenase 2 ELISA Kit is a Sandwich (quantitative) ELISA for the measurement of Human COX2/Cyclooxygenase 2 in Human in Cell/Tissue Extracts samples.

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Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

sELISA

Lab

Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

Example of human COX2/Cyclooxygenase 2 standard curve in 96-well vs. 384-well plate in 1X Cell Extraction Buffer SSW. Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Lab

Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

The concentration of COX2/Cyclooxygenase 2 was measured in human wild type and knockout A549 cell extracts based on a 250 µg/mL extract load. The mean concentration was determined to be 4,641 pg/mL in wild type extract and undetectable in knockout extract. The COX2/Cyclooxygenase 2 knockout cell line is available as ab280802.

sELISA

Lab

Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

Example of human COX2/Cyclooxygenase 2 standard curve in 1X Cell Extraction Buffer SSW. Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Lab

Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

Interpolated concentration of native COX2/Cyclooxygenase 2 was measured in duplicate at different sample concentrations. Undiluted samples are 250 µg/mL A549 cell extract, human lung tissue extract, and human colon tissue extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer SSW.

sELISA

Lab

Sandwich ELISA - Human COX2/Cyclooxygenase 2 ELISA Kit (AB325256)

Interpolated concentration of native COX2/Cyclooxygenase 2 was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 500 µg/mL lung tissue extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer SSW.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate, Tissue Extracts

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 41.4 pg/mL

Range

125 - 8000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Human COX2/Cyclooxygenase 2 ELISA Kit (ab325256) is a rapid single-wash sandwich 90-min ELISA to measure Human COX2/Cyclooxygenase 2 in Cell Extract, Tissue Extract. This SimpleStep sensitivity is 41.4 pg/ml.

How the assay works

Human COX2/Cyclooxygenase 2 ELISA Kit (ab325256) SimpleStep ELISA® employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human COX2/Cyclooxygenase 2 (ab325256) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

Save your precious samples

Our Human COX2/Cyclooxygenase 2 ELISA Kit only requires 12.5µL sample.

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

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Precision

[ { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 8.9" }, { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 5.7" } ]

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Recovery

[ { "sample": "Cell Lysate", "range": "82 - 82 %", "average": "= 82" }, { "sample": "Tissue Extracts", "range": "87 - 104 %", "average": "= 93" } ]

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What's included?

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Properties and storage information

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

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Product protocols

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Target data

Dual cyclooxygenase and peroxidase 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 (PubMed:11939906, PubMed:16373578, PubMed:19540099, PubMed:22942274, PubMed:26859324, PubMed:27226593, PubMed:7592599, PubMed:7947975, PubMed:9261177). 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 (PubMed:16373578, PubMed:22942274, PubMed:26859324, PubMed:27226593, PubMed:7592599, PubMed:7947975, PubMed:9261177). 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:16373578, PubMed:22942274, PubMed:26859324, PubMed:27226593, PubMed:7592599, PubMed:7947975, PubMed:9261177). Similarly catalyzes successive cyclooxygenation and peroxidation of dihomo-gamma-linoleate (DGLA, C20:3(n-6)) and eicosapentaenoate (EPA, C20:5(n-3)) to corresponding PGH1 and PGH3, the precursors of 1- and 3-series prostaglandins (PubMed:11939906, PubMed:19540099). In an alternative pathway of prostanoid biosynthesis, converts 2-arachidonoyl lysophopholipids to prostanoid lysophopholipids, which are then hydrolyzed by intracellular phospholipases to release free prostanoids (PubMed:27642067). Metabolizes 2-arachidonoyl glycerol yielding the glyceryl ester of PGH2, a process that can contribute to pain response (PubMed:22942274). Generates lipid mediators from n-3 and n-6 polyunsaturated fatty acids (PUFAs) via a lipoxygenase-type mechanism. Oxygenates PUFAs to hydroperoxy compounds and then reduces them to corresponding alcohols (PubMed:11034610, PubMed:11192938, PubMed:9048568, PubMed:9261177). Plays a role in the generation of resolution phase interaction products (resolvins) during both sterile and infectious inflammation (PubMed:12391014). Metabolizes docosahexaenoate (DHA, C22:6(n-3)) to 17R-HDHA, a precursor of the D-series resolvins (RvDs) (PubMed:12391014). As a component of the biosynthetic pathway of E-series resolvins (RvEs), converts eicosapentaenoate (EPA, C20:5(n-3)) primarily to 18S-HEPE that is further metabolized by ALOX5 and LTA4H to generate 18S-RvE1 and 18S-RvE2 (PubMed:21206090). In vascular endothelial cells, converts docosapentaenoate (DPA, C22:5(n-3)) to 13R-HDPA, a precursor for 13-series resolvins (RvTs) shown to activate macrophage phagocytosis during bacterial infection (PubMed:26236990). In activated leukocytes, contributes to oxygenation of hydroxyeicosatetraenoates (HETE) to diHETES (5,15-diHETE and 5,11-diHETE) (PubMed:22068350, PubMed:26282205). 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). During neuroinflammation, plays a role in neuronal secretion of specialized preresolving mediators (SPMs) 15R-lipoxin A4 that regulates phagocytic microglia (By similarity).

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