Colorimetric detection of Nitrotyrosine

Colorimetric detection of Nitrotyrosine__Colorimetric detection of Nitrotyrosine CTEP

Product Name	Nitrotyrosine ELISA Kit
Description	Colorimetric detection of Nitrotyrosine
Species Reactivity	Species Independent
Platform	Microplate
Sample Types	Cell lysates, Plasma, Serum, Urine
Detection Method	Colorimetric Assay
Assay Type	Competitive ELISA (Enzyme-linked Immunosorbent Assay)
Utility	ELISA kit used to quantitate Nitrotyrosine in samples.
Sensitivity	50 nM
Assay Range	62.5 – 8000 nM
Precision	Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate. The intra-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <10%. Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays. The inter-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <15%.
Incubation Time	1 hour
Number of Samples	38 samples in duplicate
Other Resources	, Kit Booklet , MSDS , Calculations Worksheet

Storage Temperature	4ºC
Shipping Temperature	Blue Ice
Product Type	ELISA Kits
Assay Overview	1. Prepare standard and samples in the Sample and Standard Diluent. 2. Add 50 µL of prepared standards and samples in triplicate to appropriate wells. 3. Add 50 µL of the diluted antibody preparation to the appropriate wells. 4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour. 5. Wash plate 4 times with 1X Wash Buffer. 6. Add 100 µL of TMB Substrate to each well. 7. Cover plate and develop the plate in the dark at room temperature for 30 minutes. 8. Add 100 µL of Stop Solution to each well. 9. Measure absorbance on a plate reader at 450 nm. 10. Plot the standard curve and calculate sample concentrations.
Kit Components	Component No. Item Quantity / Size SKC-126A Nitrosylated BSA Coated Plate 1 Plate SKC-126C Nitrotyrosine Standard 1 vial/110µl SKC-126F Nitrotyrosine HRP Conjugated Monoclonal Antibody 1 vial/75µl SKC-0001 Sample and Standard Diluent (Red) 1 vial/50mL SKC-0002 Nitrotyrosine Antibody Diluent (Blue) 1 vial/13mL SKC-0003 Wash Buffer Concentrate (10X) 1 vial/ 50mL SKC-0004 TMB Substrate 1 vial/13mL SKC-0005 Stop Solution 1 vial/13mL SKC-0009 Plate Cover 2
Cite This Product	Nitrotyrosine ELISA Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-126)

Alternative Names	Nitro tyrosine ELISA Kit, 3-Nitrotyrosine ELISA Kit, 3-Nitro-L-tyrosine ELISA Kit, 3 Nitrotyrosine ELISA Kit, 3-NT ELISA Kit, 3NT ELISA Kit
Research Areas	Cancer, Alzheimer_s Disease, Cell Signaling, Neurodegeneration, Neuroscience, Nitration, Oxidative Stress, Parkinson_s Disease, Post-translational Modifications
Scientific Background	Nitrotyrosine has been identified as a marker of inflammation and NO production. Nitrotyrosine is formed in presence of the active metabolite NO. Various pathways including the formation of peroxinitrite lead to nitrotyrosine production. Since nitrotyrosine is a stable end product of peroxynitrite oxidation, assessment of its plasma concentration may be useful as a marker of NO-dependent damage in vivo. Since NOX is only an indicator for enhanced NO production, protein associated nitrotyrosine might be a more suitable marker for damage induced by reactive nitrogen intermediates derived from NO. Furthermore, most proteins have a longer half life in the circulation than NOX levels. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic plaques, celiac disease, rheumatoid arthritis, chronic renal failure and septic shock. In normal plasma low, undetectable, levels of nitrotyrosine are present. Nitrosylation of the amino acid tyrosine occurs both for free tyrosine and for protein bound tyrosine.