Nitrotyrosine ELISA Kit

Analytes Available	Species Reactivity	Key Applications	Detection Methods
Nitrotyrosine	M, H, R, Po, Eq, Rb, Gp, Ht, NhP, Ca	ELISA	Chemiluminescence

Analytes Available

Species Reactivity

Key Applications

Detection Methods

Nitrotyrosine

M, H, R, Po, Eq, Rb, Gp, Ht, NhP, Ca

ELISA

Chemiluminescence

Description
Catalogue Number	17-10006
Brand Family	Upstate
Trade Name	Upstate
Description	Nitrotyrosine ELISA
Overview	Oxidative stress has been implicated in the pathogenesis of neurodegenerative disorders, cancer, and aging. Oxidative stress occurs as a response to increased oxidants, decreased anti-oxidants, or failure to repair oxidative damage induced by reactive oxygen species (ROS). ROS are free radicals, reactive molecules containing oxygen, or molecules containing oxygen that generate free radicals. ROS include nitric oxide (NO), superoxide (O₂^-), peroxynitrite (ONOO^-), and hydroxyl radical (OH^-). ROS are cleared from cells by the action of superoxide dismutase (SOD), catalase and peroxidases. NO, produced by nitric oxide synthases (NOS), is a critical signaling molecule involved in many physiological processes including muscle contraction, vasodilation, neuronal transmission, and immune responses. Some of these effects are mediated through reaction with and activation of guanylate cyclase (GC), resulting in increased cGMP production. With a half-life of seconds, NO readily diffuses through the cytosol and cell membrane, allowing it to act inside or adjacent to the cell in which it was produced. ROS may interact to form agents that modify cellular proteins. Superoxide, produced by NADPH oxidase (phox), reacts with NO to form peroxynitrite. Intracellular peroxynitrite can modify proteins by interacting with and nitrating tyrosine residues to form 3-nitrotyrosine. Tyrosine nitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. Mn-SOD, Ca⁺⁺-ATPase), or have no discernable effect (e.g. p53, VASP, α-Synuclein) on the activity of a particular protein. Upstate's new Nitrotyrosine Assay Kit, Chemiluminescence Detection (Cat. # 17-10006) is a competitive ELISA for the quantitation of tyrosine nitration. The kit includes all reagents needed for measurement of tyrosine nitration, including a pre-coated 96-well plates, Nitrotyrosine peptide standard, a Nitrotyrosine antibody, LumiGLO® chemiluminescent detection substrate, and wash buffers. The assay has a wide dynamic range and high precision, making this assay a valuable new tool for the study of tyrosine nitration.
Materials Required but Not Delivered	1. Multi-channel or repeating pipettes 2. Plate shaker (optional) 3. Pipettors and tips capable of accurately measuring 1-1000 µL 4. Graduated serological pipettes 5. 96-well microtiter plate reader with luminescence readout. 6. Graphing software for plotting data or graph paper for manual plotting of data 7. Microcentrifuge tubes for standard and sample dilutions 8. Mechanical vortex 9. 1 liter container 10. Distilled or deionized water
Background Information	Oxidative stress has been implicated in the pathogenesis of neurodegenerative disorders, cancer and aging. Oxidative stress occurs as a response to increased oxidants, decreased antioxidants, or failure to repair oxidative damage induced by reactive oxygen species (ROS). ROS are free radicals, reactive molecules containing oxygen, or molecules containing oxygen that generate free radicals. ROS include nitric oxide (NO), superoxide (O2-), peroxynitrite (ONOO-), and hydroxyl radical (OH-). ROS are cleared from cells by the action of superoxide dismutase (SOD), catalase and peroxidases. NO, produced by nitric oxide synthases (NOS), is a critical signaling molecule involved in many physiological processes including muscle contraction, vasodilation, neuronal transmission, and immune responses. Some of these effects are mediated through reaction with and activation of guanylate cyclase (GC), resulting in increased cGMP production. With a half-life of seconds, NO readily diffuses through the cytosol and cell membrane, allowing it to act inside or adjacent to the cell in which it was produced. ROS may interact to form agents that modify cellular proteins. Superoxide, produced by NADPH oxidase (phox), reacts with NO to form peroxynitrite. Intracellular peroxynitrite can modify proteins by interacting with and nitrating tyrosine residues to form 3-nitrotyrosine. Tyrosine nitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. Mn-SOD, Ca++-ATPase), or have no discernable effect (e.g. p53, VASP, α-Synuclein) on the activity of a particular protein.

Description

Catalogue Number

17-10006

Brand Family

Upstate

Trade Name

Upstate

Description

Nitrotyrosine ELISA

Overview

Oxidative stress has been implicated in the pathogenesis of neurodegenerative disorders, cancer, and aging. Oxidative stress occurs as a response to increased oxidants, decreased anti-oxidants, or failure to repair oxidative damage induced by reactive oxygen species (ROS). ROS are free radicals, reactive molecules containing oxygen, or molecules containing oxygen that generate free radicals. ROS include nitric oxide (NO), superoxide (O₂^-), peroxynitrite (ONOO^-), and hydroxyl radical (OH^-). ROS are cleared from cells by the action of superoxide dismutase (SOD), catalase and peroxidases.

NO, produced by nitric oxide synthases (NOS), is a critical signaling molecule involved in many physiological processes including muscle contraction, vasodilation, neuronal transmission, and immune responses. Some of these effects are mediated through reaction with and activation of guanylate cyclase (GC), resulting in increased cGMP production. With a half-life of seconds, NO readily diffuses through the cytosol and cell membrane, allowing it to act inside or adjacent to the cell in which it was produced.

ROS may interact to form agents that modify cellular proteins. Superoxide, produced by NADPH oxidase (phox), reacts with NO to form peroxynitrite. Intracellular peroxynitrite can modify proteins by interacting with and nitrating tyrosine residues to form 3-nitrotyrosine. Tyrosine nitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. Mn-SOD, Ca⁺⁺-ATPase), or have no discernable effect (e.g. p53, VASP, α-Synuclein) on the activity of a particular protein.

Upstate's new Nitrotyrosine Assay Kit, Chemiluminescence Detection (Cat. # 17-10006) is a competitive ELISA for the quantitation of tyrosine nitration. The kit includes all reagents needed for measurement of tyrosine nitration, including a pre-coated 96-well plates, Nitrotyrosine peptide standard, a Nitrotyrosine antibody, LumiGLO® chemiluminescent detection substrate, and wash buffers.

The assay has a wide dynamic range and high precision, making this assay a valuable new tool for the study of tyrosine nitration.

Materials Required but Not Delivered

1. Multi-channel or repeating pipettes
2. Plate shaker (optional)
3. Pipettors and tips capable of accurately measuring 1-1000 µL
4. Graduated serological pipettes
5. 96-well microtiter plate reader with luminescence readout.
6. Graphing software for plotting data or graph paper for manual plotting of data
7. Microcentrifuge tubes for standard and sample dilutions
8. Mechanical vortex
9. 1 liter container
10. Distilled or deionized water

Background Information

Oxidative stress has been implicated in the pathogenesis of neurodegenerative disorders, cancer and aging. Oxidative stress occurs as a response to increased oxidants, decreased antioxidants, or failure to repair oxidative damage induced by reactive oxygen species (ROS). ROS are free radicals, reactive molecules containing oxygen, or molecules containing oxygen that generate free radicals. ROS include nitric oxide (NO), superoxide (O2-), peroxynitrite (ONOO-), and hydroxyl radical (OH-). ROS are cleared from cells by the action of superoxide dismutase (SOD), catalase and peroxidases.
NO, produced by nitric oxide synthases (NOS), is a critical signaling molecule involved in many physiological processes including muscle contraction, vasodilation, neuronal transmission, and immune responses. Some of these effects are mediated through reaction with and activation of guanylate cyclase (GC), resulting in increased cGMP production. With a half-life of seconds, NO readily diffuses through the cytosol and cell membrane, allowing it to act inside or adjacent to the cell in which it was produced.
ROS may interact to form agents that modify cellular proteins. Superoxide, produced by NADPH oxidase (phox), reacts with NO to form peroxynitrite. Intracellular peroxynitrite can modify proteins by interacting with and nitrating tyrosine residues to form 3-nitrotyrosine. Tyrosine nitration may increase (e.g. sGC, Src, PI3K, Akt), decrease (e.g. Mn-SOD, Ca++-ATPase), or have no discernable effect (e.g. p53, VASP, α-Synuclein) on the activity of a particular protein.

Product Information
Components	1. Streptavidin Capture Plate: (Part No. CS204971) One pre-coated 96-stripwell immunoplate sealed in a foil pouch. 2. U-Bottom Plate: (Part No. CS205781) One clear non-binding U-bottom plate. 3. TBS, 20X: (Part No. 20-190) One bottle (50 mL) of 50X TBS. 4. 20% Tween® 20: (Part No. 20-246) One bottle (3 mL) of 20% Tween® 20 (v/v). 5. LumiGlo Chemiluminescent Substrate Reagent A: (Part No. 20-212C) One bottle (10 mL) of substrate. 6. LumiGlo Chemiluminescent Substrate Reagent B: (Part No. 20-212D) One bottle (10 mL) of substrate. 7. Plate Covers: Two plate covers. 8. Nitrotyrosine Peptide Standard: (Part No. CS204980). One vial containing (200 µL) of Nitrotyrosine Peptide Standard Stock at 4.8mM. 9. 1000X Biotinylated Nitrotyrosine Peptide: (Part No. CS204990). One vial containing (15 µL) of Biotinylated Nitortyrosine Peptide Stock at 450µM. 10. 2000X Anti-Nitrotyrosine Detection Antibody: (Part No. CS204982) One vial (10 µL) of 2000x anti-Nitrotyrosine detection antibody. 11. Gt X Rb, HRP Secondary Antibody: (Part No. 90276) One vial (30 µL) of 2000X goat anti-rabbit HRP conjugate. 12. 30% BSA: (Part No. CS205797) One vial containing 1mL of 30% BSA.
Detection method	Chemiluminescence

Product Information

Components

1. Streptavidin Capture Plate: (Part No. CS204971) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.
2. U-Bottom Plate: (Part No. CS205781) One clear non-binding U-bottom plate.
3. TBS, 20X: (Part No. 20-190) One bottle (50 mL) of 50X TBS.
4. 20% Tween® 20: (Part No. 20-246) One bottle (3 mL) of 20% Tween® 20 (v/v).
5. LumiGlo Chemiluminescent Substrate Reagent A: (Part No. 20-212C) One bottle (10 mL) of substrate.
6. LumiGlo Chemiluminescent Substrate Reagent B: (Part No. 20-212D) One bottle (10 mL) of substrate.
7. Plate Covers: Two plate covers.
8. Nitrotyrosine Peptide Standard: (Part No. CS204980). One vial containing (200 µL) of Nitrotyrosine Peptide Standard Stock at 4.8mM.
9. 1000X Biotinylated Nitrotyrosine Peptide: (Part No. CS204990). One vial containing (15 µL) of Biotinylated Nitortyrosine Peptide Stock at 450µM.
10. 2000X Anti-Nitrotyrosine Detection Antibody: (Part No. CS204982) One vial (10 µL) of 2000x anti-Nitrotyrosine detection antibody.
11. Gt X Rb, HRP Secondary Antibody: (Part No. 90276) One vial (30 µL) of 2000X goat anti-rabbit HRP conjugate.
12. 30% BSA: (Part No. CS205797) One vial containing 1mL of 30% BSA.

Detection method

Chemiluminescence

Applications
Application	This Nitrotyrosine ELISA is used to measure & quantify Nitrotyrosine levels in Neuroscience & Toxicity research.
Key Applications	ELISA

Applications

Application

This Nitrotyrosine ELISA is used to measure & quantify Nitrotyrosine levels in Neuroscience & Toxicity research.

Key Applications

ELISA

Biological Information
Species Reactivity	Mouse Human Rat Pig Horse Rabbit Guinea Pig Hamster Non-human Primate Canine
Analytes Available	Nitrotyrosine

Biological Information

Species Reactivity

Mouse
Human
Rat
Pig
Horse
Rabbit
Guinea Pig
Hamster
Non-human Primate
Canine

Analytes Available

Nitrotyrosine

Physicochemical Information
Sensitivity	0.1 μM
Standard Curve Range	0.1 μM - 1600 μM

Physicochemical Information

Sensitivity

0.1 μM

Standard Curve Range

0.1 μM - 1600 μM

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Maintain the unopened kit (17-10006-1) at 2-8ºC until expiration date. Maintain the unopened kit (17-10006-2) at -20ºC until expiration date. Avoid repeated freeze/thaw cycles, aliquot if necessary.

Storage and Shipping Information

Storage Conditions

Maintain the unopened kit (17-10006-1) at 2-8ºC until expiration date. Maintain the unopened kit (17-10006-2) at -20ºC until expiration date. Avoid repeated freeze/thaw cycles, aliquot if necessary.

Packaging Information
Material Size	1 kit

Packaging Information

Material Size

1 kit

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Nitrotyrosine ELISA Kit

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