Oxidative stress markers

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Oxidative stress markers

Oxidative stress represents an imbalance between reactive oxygen species (ROS) and cellular mechanisms for detoxifying the reactive intermediates or for repairing the resulting damage. Disturbances in the normal state can cause toxic effects through the production of peroxides and free radicals that damage all components of the cell, including proteins, lipids, and DNA.

Nowadays, oxidative stress may involve cell perturbations leading to various diseases like atherosclerosis, heart failure, myocardial infarction, Alzheimer’s disease, schizophrenia…

Oxidative Stress may also be important in the prevention of aging by induction of a process named mitohormesis. ROS can be beneficial, as they are used by the immune system as a way to attack and kill pathogens.

Advanced BioDesign offers you a wide range of products for monitoring oxidative stress in cells or tissues by following specific marker of DNA oxidation, lipid peroxidation, protein oxidation.

Selection Guide for Oxidative Stress Assays by Sample Type

Cells Urine Serum Tissue New 8-OHdG Check ELISA kit

02-0101

Measuring Range:

0.5 - 200 ng/mL X X X Assay time: approx 3.5 hours.

Highly Sensitive 8-OHdG Check ELISA kit 02-0102

Measuring Range:

0.125 - 10 ng/mL X X X X Assay time: overnight

Anti 8-OHdG monoclonal antibody

02-0201 X X

Suitable for immunohistochemistry (IHC) and ELISA methods

Anti Tymidine Glycol (TG) monoclonal antibody

02-0501 X X Suitable for IHC

Hexanoyl-Lysine (HEL) ELISA kit 02-0701

Measuring Range:

2 - 700 nmol/L X X X X

New biomarkers for early stage of lipid oxidation.

Anti Hexanoyl-Lysine (HEL) monoclonal antibody

02-0702 X X

Suitable for IHC, Western Blot (WB) and ELISA

Anti 4-HNE monoclonal antibody

02-0301 X X Suitable for IHC and WB

Anti acrolein (ACR) monoclonal antibody

02-0901 X X Suitable for IHC, WB and ELISA

Anti malondialdehyde (MDA) monoclonal antibody

Anti 4-hydroxy-2-hexenal (4-HHE) monoclonal antibody

Anti crotonaldehyde (CRA) monoclonal antibody

Anti methyglyoxal (MG) monoclonal antibody

Anti 7-ketocholesterol (7-KC) monoclonal antibody

Anti Di bromo tyrosine (DiBrY) monoclonal antibody

Anti-DiTyrosine (DT) monoclonal antibody

DNA Damage Markers

Lipid Peroxydation Markers

Protein Oxidation Markers

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DNA Oxidation Damages Markers

Oxidative DNA damage is an inevitable consequence of cellular metabolism, with a propensity for increased levels following toxic insult. Although more than 20 base lesions have been identified, only a fraction of these have received appreciable study.

8-OHdG ELISA kit (8-hydroxy-2’-deoxyguanosine)

8-hydroxy-2′-deoxyguanosine (8-OHdG) is a product of oxidatively damaged DNA formed by hydroxy radical, singlet oxygen and direct photodynamic action. 8-OHdG can be detected in tissue, serum, urine and other biomaterials. 8-OHdG Check is a competitive enzyme-linked immunosorbent assay (ELISA) utilising monoclonal antibody (clone N45.1) which is highly specific for DNA damage,and does not cross react with RNA oxidation products such as 8-hydroxy-guanine and 8-hydroxy-guanosine. This product is suitable for detection of 8-OHdG in urine and other biomaterials from human and animals.

1. High specificity & high sensitivity: Cross reactivity was checked for 19 analogues. 2. Easy operation & speedy:There is no need for expensive equipment and sample

pretreatment. Required equipments are: pipettes, incubator and micro plate reader with 450nm filter. Assays can be completed within 3.5 hours (New 8OHdG Check). Reference: S.Saito, et al.: Res. Commun. Mol. Pathol. Pharmacol. 107 (1&2), p39-44 (2000)

Product reference:

 02-0101: 8-OHdG Check ELISA kit

 02-0102: Highly Sensitive 8-OHdG Check ELISA kit

 02-0103: 8-OHdG Check ELISA Trial Package (32wells) Anti 8-OHdG monoclonal antibody (clone N45.1)

Highly Specific for 8-OHdG

Cross reactivity was checked for 19 analogues:

7-methyl-G, 6-SH-G, 8-bromo-G, dA, dC, dT, dI, dU, dG, O6-methyl-dG, 8-OHdA, guanine (Gua), O6-methyl-Gua, 8-OHGua, uric acid, Urea, creatine, creatinine} demonstrate no cross-reactivity.

Only 8-sulfhydryl-G and 8-OHG demonstrate minimal cross-reactivity (less than 1%).

Product reference:

 02-0201: Anti 8-OHdG monoclonal antibody (100µg)

 02-0202: Anti 8-OHdG monoclonal antibody (20µg)

Anti Thymidineglycol (TG) monoclonal antibody

Thymidineglycol (TG) is one of the major oxidation products of DNA. Thymidine (T) can be damaged by oxidative stress such as radiation and energy metabolism. Two different pathways to form TG have been suggested. Deoxythymidine in DNA is directly oxidised by hydroxy radical, to form TG. TG can be also formed through an intermediate thymidine chlorohydrin, which is derived from hypochlorous acid (HOCl) from neutrophil myeloperoxidase. Thymidineglycol is derived from DNA, not from RNA. TG is the oxidative stress marker specific for DNA damage.

Reference: Ashis K et al.: Proc.Natl.Acad.Sci.Vol.86, p76777681 (1989) Product reference:

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Lipid peroxidation markers

Lipid peroxidation refers to the oxidative degradation of lipids. It is the process in which free radicals "steal" electrons from the lipids in cell membranes, resulting in cell damage. This process proceeds by a free radical chain reaction mechanism. It most often affects polyunsaturated fatty acids, because they contain multiple double bonds in between which lie methylene -CH2- groups that possess especially reactive hydrogens

Anti 4-HNE monoclonal antibody (HNE-J2)

Membrane lipids are one of the targets of reactive oxygen species (ROS). 4-hydroxy-2-nonenal (4-HNE) is a major membrane lipid peroxidation product. Monoclonal antibody against 4-HNE (clone HNE-J2) is highly specific for 4-HNE-His/Lys/Cys adducts.

Product reference

 02-0301: Anti 4-HNE monoclonal antibody (100µg)

 02-0302: Anti 4-HNE monoclonal antibody (20µg) Anti 4-HHE (4-hydroxy-2-hexenal) monoclonal antibody

4-hydroxy-2-alkenal is one of the major lipid peroxidation products, and shows many biological effects such as high toxicity to cells. Among them, 4-hydroxy-2-hexenal (HHE) is an aldehyde formed during peroxidation of n-3 fatty acids such as docosahexaenoic acid. HHE is a highly reactive aldehyde that reacts with histidine residues of proteins to form Michael-addition type adducts. This antibody is specific for HHE-histidine Michael adduct (HHE-His) and permits HHE-His to be detected in tissue samples.

 02-1001: Anti 4-hydroxy-2-hexenal (4-HHE) monoclonal antibody (30µg) Hexanoyllysine adduct (HEL) ELISA kit & antibody

A new biomarker of lipid peroxidation.

Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in some diseases, pertubation of cell functions and aging. Aldehydes such as malondi-aldehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) have been reported as one of the advanced lipid peroxidation products. But recently in the earlier stage of lipid peroxidation, 13-hydroperoxyoctadecanoic acid (13-HPODE) is found to be covalently bound to proteins1). Hexanoyl-Lysine adduct (HEL) is a novel lipid hydroperoxide-modified lysine residues. HEL is formed by oxidative modification of oxidized omega-6 fatty acids such as linoleic acid or arachidonic acid. HEL may be a useful biomarker for initial stage of lipid peroxidation.Monoclonal antibodies and an ELISA kit have been developped, and HEL can be detected in oxidatively modified LDL, in human atherosclerotic lesions, human urine and serum. It is also reported that HEL is formed in rat muscle during exercise, and the formation is prohibited by antioxidants such as flavonoids.

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 02-0701: Hexanoyllysine(HEL) ELISA kit - suitable for urine and serum samples.

 02-0702: Anti Hexanoyl-Lys(HEL) monoclonal antibody (clone 5F12) (20µg) – for IHC & WB

Anti 7-ketocholesterol monoclonal antibody

Cholesterol oxidation products, especially 7-Ketocholesterol (7KC), have been the focus of much attention because they are present in human atherosclerotic plaques and display a wide range of atherogenic properties in vitro and to some extent in vivo.

This antibody is specific for 7KC and especially in tissue samples. Frozen tissue sample is recommended for immunohistochemistry. Product reference

 02-1301: Anti 7-ketocholesterol (7-KC) monoclonal antibody (20µg)

Anti Crotonaldehyde monoclonal antibody

Crotonaldehyde (CRA) is a representative carcinogenic aldehyde formed endogenously through lipid peroxidation. CRA is a highly reactive aldehyde that reacts with lysine residues in proteins. The reaction between CRA and lysine residue leads to the formation of numerous adducts.

This antibody is specific for the CRA-modified protein. Product reference

 02-1101: Anti-Crotonaldehyde (CRA) monoclonal antibody (30µg) Anti-Malondialdehyde monoclonal antibody

Malondialdehyde (MDA) is one of the major aldehydes derived from lipid peroxidation. MDA is a highly reactive aldehyde that reacts with lysine residues in proteins. The reaction of MDA with lysine residues leads to the formation of many types of adducts, such as dihydropyridine-lysine (DHP-lysine) types of derivatives.

This monoclonal antibody is specific for the MDA-modified protein, especially DHP-lysine type derivative.

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Protein oxidation damages

Anti Methylglyoxal monoclonal antibody

Methylglyoxal (MG), an endogenous metabolite that increases in diabetes and is a common intermediate in the Maillard reaction (glycation), reacts with proteins and forms advanced glycation end products (AGE). MG reacts with arginine residues in proteins and forms numerous numbers of adducts, such as argpyrimidine. This antibody is specific for argpyrimidine: N-(5-hydroxy-4,6-dimethylpyrimidine-2-yl)-L-ornithine.

 02-1201: Anti-methylglyoxal monoclonal antibody Anti Dityrosine monoclonal antibody

Dityrosine a novel marker for protein oxidation

Tyrosine is one of the major targets of protein oxidation, and until today various tyrosine derivatives such as nitrotyrosine, dityrosine (DT) and halogenated tyrosine depending on the type of free radicals. DT is a tyrosine dimer derived from tyrosyl radicals which is formed by reactive oxygen species (ROS), metal-catalyzed oxidation, ultraviolet irradiation, and peroxidases. Dityrosine has been found in atherosclerotic lesions, and lipofuscin of pyramidal neurons of aged human brains. Dityrosine is one of the specific biomarkers for protein oxidation.

 02-0601: Anti-DiTyrosine (DT) monoclonal antibody Anti Dibromo-Tyrosine monoclonal antibody

Neutrophils and eosinophils play an important role in the defence system against microbial infection. Myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are known to catalyze formation of hypochlorous acid (HOCl) and hypobromous acid (HOBr). These reactive intermediates may react with proteins, lipids and nucleotides, and it has been reported to form tyrosine halogenation products such as dibromotyrosine (DiBrY). DiBrY is a Br-modified tyrosine at 3- and 5- positions, which is one of the major oxidative products derived from neutrophil myeloperoxidase.

 02-0401: Anti-Dibromo-Tyrosine (DiBrY) monoclonal antibody (20µg)

Advanced BioDesign

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