Nitrotyrosine ELISA Kit: 96 Tests

NWLSS | Northwest Life Science Specialties, LLC.
nwk-ntr0101
NWK-NTR01-01 $645.00 each

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NWK-NTR01-01
The NWLSS™ Nitrotyrosine ELISA is recommended for the detection of nitrosylated proteins in plasma or serum samples. Excellent tool for peroxynitrate related studies and recommended as a biomarker of protein related oxidative/nitrosative stress related augmentation.

Introduction

Nitrate (NO3) and nitrite (NO2) production are typically used as markers of nitric oxide production (NO) production. These analytes by themselves fail to address the eventual fate of NO or the possible adverse effects associated with its excess production and reaction with free radical species in vivo. Active NO metabolites can react with superoxide to form peroxynitrite (ONOO-) a powerful oxidant and nitrating agent. Subsequent reaction of peroxynitrite with proteins results in nitrotyrosine formation. As a stable end product of peroxynitrite mediated oxidation/nitration, Nitrotyrosine can be used as a surrogate index of NO dependent damage in vivo and has been associated with multiple disease states.

Test Principle

The NWLSS™ Nitrotyrosine ELISA test is a simple "sandwich" ELISA using a plate bound capture antibody (anti nitrated KLH) to nitrotyrosine and a biotinylated secondary tracer antibody. Addition of streptavidin-peroxidase followed by tetramethylbenzidine (TMB) facilitates color development directly proportional to the nitrotyrosine present in the sample. The reaction is stopped using a citric acid solution and the assay is read on a plate reader at 450 nm.

View Assay Protocol

Catalog Number: NWK-NTR01-01
Methodology: 96 wells presented as 1 plate sectionable as 8 well individual strips
Intended Use Quantitative measurement of nitrotyrosine protein adduct in biological samples. The NWLSS™ product allows quantification of oxidative/nitrosative stress effect on proteins and peroxynitrite formation in biological systems.
Sample Requirements: Plasma, serum and other biological samples such as tissue samples.
Specificity: Nitrosylated protein adducts
Sensitivity: 2 nM in assay
Storage and Stability: 1 year from manufacture date when stored at specified temperature
Kit Contents: Precoted Microwell strips and strip frames
Wash Buffer
Dilution Buffer
Nitrotyrosine Standard
Biotinylated anti nitrotyrosine tracer
Streptavidin Peroxidase
TMB Substrate
Stop Solution

Does the NWK-NTR01 ELISA detect free nitrotyrosine or nitrosylated protein adducts?

The NWK-NTR01 ELISA recognizes nitrotyrosine-protein adducts. {sliders Question: How should tissues be homogenized for assay using the NWK-NTR01 kit?} Answer: We normally recommend that tissue samples re homogenized in normal PBS, pH 7.0. Homogenization can be performed with a dounce style or Potter Elvehjem style tissue grinder or similar apparatus. Tissue homogenates should be centrifuged and the clarified supernatant tested for nitrotyrosine. Several references wherein other researchers have used the NWK-NTR01 kit to test tissue samples can be found in the citations references for this product.

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