OxyIHC™ Oxidative Stress Detection Kit

Key Applications	Detection Methods
IHC	Chromogenic

Key Applications

Detection Methods

IHC

Chromogenic

Description
Catalogue Number	S7450
Trade Name	OxyIHC
Description	OxyIHC™ Oxidative Stress Detection Kit
Overview	The OxyIHC™ Oxidative Stress Detection Kit (S7450) contains the chemical and immunological reagents necessary to detect protein oxidation in various tissues from a variety of organs and animal species. The OxyIHC test method involves chemical derivatization of protein carbonyl groups with 2,4- dinitrophenylhydrazine (DNPH). This chemical reaction results in proteins being covalently coupled to DNP at their carbonyl sites. The DNP-derivatized proteins are then detected using an antibody that specifically binds to the DNP moiety. Subsequent incubation with biotin conjugated secondary antibody, streptavidin conjugated HRP, and development using a 3,3’ diaminobenzidine (DAB) staining allows immunohistochemical detection of protein oxidation.
Background Information	Reactive oxygen species (ROS) have been implicated in pathological processes including cancer, apoptosis, aging, neurodegenerative diseases, chronic inflammatory diseases, pulmonary diseases, and cardiovascular diseases (for reviews, see ref. 1-4). Living organisms are continually exposed to potentially harmful oxygen free radicals that are generated thru normal cellular functions as well as from environmental factors (1, 5-7). Yet, these reactive molecules are mitigated by the presence of antioxidants and therefore organisms exist with a delicate balance between oxidants and antioxidants (7-9). During times of oxidative stress there is an imbalance in favor of the oxidants. The end result of which is that free radicals are able to attack and modify sub-cellular components including nucleic acids, lipids and proteins (7, 10-12). Proteins are one of the major targets of reactive species. Oxidation of proteins modifies the side chains of methionine, histidine, and tyrosine and forms cysteine disulfide bonds (16-19). Metal catalyzed oxidation of proteins introduces carbonyl groups (aldehydes and ketones) at lysine, arginine, proline or threonine residues in a site-specific manner (16, 20-22). The oxidative modification of proteins can modulate biochemical characteristics of proteins such as enzymatic activity (21-23), DNA binding activities of transcription factors (24-26), and the susceptibility to proteolytic degradation (12, 25-28). While a relationship between protein oxidation and aging has been suggested (29-31), little is known about the importance of oxidative modification of individual proteins in the pathophysiology of free radical mediated processes. Not surprisingly, carbonyl modification of proteins has become a key biomarker for the identification of oxidative stress.

Description

Catalogue Number

S7450

Trade Name

OxyIHC

Description

OxyIHC™ Oxidative Stress Detection Kit

Overview

The OxyIHC™ Oxidative Stress Detection Kit (S7450) contains the chemical and immunological reagents
necessary to detect protein oxidation in various tissues from a variety of organs and animal species. The
OxyIHC test method involves chemical derivatization of protein carbonyl groups with 2,4-
dinitrophenylhydrazine (DNPH). This chemical reaction results in proteins being covalently coupled to DNP
at their carbonyl sites. The DNP-derivatized proteins are then detected using an antibody that specifically
binds to the DNP moiety. Subsequent incubation with biotin conjugated secondary antibody, streptavidin
conjugated HRP, and development using a 3,3’ diaminobenzidine (DAB) staining allows
immunohistochemical detection of protein oxidation.

Background Information

Reactive oxygen species (ROS) have been implicated in pathological processes including cancer,
apoptosis, aging, neurodegenerative diseases, chronic inflammatory diseases, pulmonary diseases, and
cardiovascular diseases (for reviews, see ref. 1-4). Living organisms are continually exposed to potentially
harmful oxygen free radicals that are generated thru normal cellular functions as well as from environmental
factors (1, 5-7). Yet, these reactive molecules are mitigated by the presence of antioxidants and therefore
organisms exist with a delicate balance between oxidants and antioxidants (7-9). During times of oxidative
stress there is an imbalance in favor of the oxidants. The end result of which is that free radicals are able to
attack and modify sub-cellular components including nucleic acids, lipids and proteins (7, 10-12).
Proteins are one of the major targets of reactive species. Oxidation of proteins modifies the side chains of
methionine, histidine, and tyrosine and forms cysteine disulfide bonds (16-19). Metal catalyzed oxidation of
proteins introduces carbonyl groups (aldehydes and ketones) at lysine, arginine, proline or threonine
residues in a site-specific manner (16, 20-22). The oxidative modification of proteins can modulate
biochemical characteristics of proteins such as enzymatic activity (21-23), DNA binding activities of
transcription factors (24-26), and the susceptibility to proteolytic degradation (12, 25-28). While a
relationship between protein oxidation and aging has been suggested (29-31), little is known about the
importance of oxidative modification of individual proteins in the pathophysiology of free radical mediated
processes. Not surprisingly, carbonyl modification of proteins has become a key biomarker for the
identification of oxidative stress.

Product Information
Components	10X Wash Buffer, One 25 mL bottle 10X Antigen Retrieval Buffer, One 1.25 mL bottle 5X Blocking Buffer, One 2.5 mL bottle Primary Antibody, One 125 mL vial DAB-A, One 500 mL vial DAB-B, One 12 mL bottle Antibody Diluent, One ready to use 12.5 mL bottle Biotinylated Secondary Antibody, One ready to use 12.5 mL bottle Streptavidin conjugated HRP, One ready to use 12.5 mL bottle Hematoxylin, One ready to use 12.5 mL bottle DNPH Solution, One ready to use 12.5 mL bottle Derivatization Control Solution, One ready to use 12.5 mL bottle
Detection method	Chromogenic

Product Information

Components

10X Wash Buffer, One 25 mL bottle
10X Antigen Retrieval Buffer, One 1.25 mL bottle
5X Blocking Buffer, One 2.5 mL bottle
Primary Antibody, One 125 mL vial
DAB-A, One 500 mL vial
DAB-B, One 12 mL bottle
Antibody Diluent, One ready to use 12.5 mL bottle
Biotinylated Secondary Antibody, One ready to use 12.5 mL bottle
Streptavidin conjugated HRP, One ready to use 12.5 mL bottle
Hematoxylin, One ready to use 12.5 mL bottle
DNPH Solution, One ready to use 12.5 mL bottle
Derivatization Control Solution, One ready to use 12.5 mL bottle

Detection method

Chromogenic

Applications
Application	The OxyIHC Oxidative Stress Detection Kit contains the chemical & immunological reagents necessary to detect protein oxidation in various tissues from a variety of organs & animal species.
Key Applications	Immunohistochemistry

Applications

Application

The OxyIHC Oxidative Stress Detection Kit contains the chemical & immunological reagents necessary to detect protein oxidation in various tissues from a variety of organs & animal species.

Key Applications

Immunohistochemistry

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	The OxyIHC kit components should be stored at 2-8ºC. Kit components are stable for 6 months from the date of receipt if stored and handled properly.

Storage and Shipping Information

Storage Conditions

The OxyIHC kit components should be stored at 2-8ºC. Kit components are stable for 6 months from the
date of receipt if stored and handled properly.

Packaging Information
Material Size	50 Reactions

Packaging Information

Material Size

50 Reactions

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

OxyIHC™ Oxidative Stress Detection Kit

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