AB316905

3-Nitrotyrosine ELISA Kit - Intracellular

SimpleStep
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3-Nitrotyrosine ELISA Kit - Intracellular is a single-wash 90-min Simplestep used to quantify 3-Nitrotyrosine with a sensitivity of 40.3 ng/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Competitive ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders

4 Images

Competitive ELISA - 3-Nitrotyrosine ELISA Kit - Intracellular (AB316905)

cELISA

Supplier Data

Competitive ELISA - 3-Nitrotyrosine ELISA Kit - Intracellular (AB316905)

Recombinant 3-Nitrotyrosinewas spiked into the following samples and diluted in a 2-fold dilution series in 1X Cell Extraction Buffer PTR. Undiluted samples are 100 µg/mL HepG2 cell extract, PC-12 cell extract, and human heart tissue extract. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

cELISA

Supplier Data

Competitive ELISA - 3-Nitrotyrosine ELISA Kit - Intracellular (AB316905)

Comparison of native 3-Nitrotyrosine concentration in treated versus control cell extract. HepG2 cells were treated with 10 mM peroxynitrite diluted in PBS for 10 minutes, scraped from the plate and then extracted with 1X Cell Extraction Buffer PTR. Mock control HepG2 cells accompanied cell treatment. Concentrations of 3-Nitrotyrosine were measured in duplicate and interpolated against the standard curve. Results show the mean 3-Nitrotyrosine concentration in control cell extract was 98 ng/mL compared to 1738 ng/mL in peroxynitrite-treated sample.

cELISA

Supplier Data

Competitive ELISA - 3-Nitrotyrosine ELISA Kit - Intracellular (AB316905)

Interpolated concentration of native 3-Nitrotyrosine was measured in duplicate at different sample concentrations. Undiluted sample is peroxynitrite-treated HepG2 cell extract 2.5 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.

cELISA

Supplier Data

Competitive ELISA - 3-Nitrotyrosine ELISA Kit - Intracellular (AB316905)

Example of 3-Nitrotyrosine standard curve. OD (450 nm) values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate, Tissue Extracts

Assay type

Competitive

Sensitivity

= 40.3 ng/mL

Range

62.5 - 4000 ng/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": { "cELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

3-Nitrotyrosine SimpleStep ELISA® kit is a single-wash 90 min competitive ELISA designed for the quantitative measurement of 3-Nitrotyrosine in Cell Lysate and Tissue Extract. Quantitate 3-Nitrotyrosine with 40.3 ng/mL sensitivity.

SimpleStep ELISA® technology employs capture conjugates conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to competitive ELISA allows the formation of the antibody-analyte complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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Precision

[ { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "6.2" }, { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.9" } ]

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Recovery

[ { "sample": "HepG2 Extract", "range": "92 - 105 %", "average": "= 99" }, { "sample": "Human Heart Extract", "range": "80 - 83 %", "average": "= 81" }, { "sample": "PC12 Extract", "range": "89 - 106 %", "average": "= 105" } ]

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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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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

3-Nitrotyrosine forms when peroxynitrite interacts with tyrosine residues in proteins. It is a marker of nitrosative stress and oxidative damage with the molecular mass of the modified residue remaining similar to that of tyrosine. This molecule is commonly found in conditions with high oxidative stress such as inflammation and is present in tissues with significant reactive nitrogen species. 3-Nitrotyrosine detection is facilitated by various techniques including nitrotyrosine staining and plays a notable role in the study of oxidative stress markers.

Biological function summary

Modification of tyrosine can affect protein function and cell signaling. This post-translational modification can hinder protein function or disrupt protein-protein interactions altering cellular processes. Nitrotyrosine often forms part of the cellular response to oxidative damage contributing to complex antioxidant defenses. The presence of nitrotyrosine indicates a cellular environment stressed by reactive nitrogen species linking it to pathophysiological processes in cells undergoing such stress.

Pathways

3-nitrotyrosine acts in oxidative stress and cell signaling routes. It integrates into pathways related to the cellular stress response particularly the antioxidant defense mechanisms. Proteins such as superoxide dismutase (SOD) may indirectly relate to nitrotyrosine since they mitigate reactive species trying to maintain redox balance. The presence of nitrotyrosine can indicate excess stress on these systems reflecting shifts in signaling that affect cell survival.

High levels of 3-nitrotyrosine associate with cardiovascular diseases and neurodegenerative disorders. In diseases such as atherosclerosis and Alzheimer's 3-nitrotyrosine presence indicates elevated oxidative stress. Through these conditions it relates to proteins like amyloid-beta in Alzheimer's disease or low-density lipoprotein (LDL) in cardiovascular disease. These associations emphasize the potential of 3-nitrotyrosine as a biomarker for oxidative damage in stress-related diseases.

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Product protocols

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Target data

See full target information 3-Nitrotyrosine

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websiteProtocolBooklet

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Product promise

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