JaICA
Japan Institute for the Control of Aging

Oxidative Stress Markers
CAT20080222E① 8­OHdG ELISA kit (8­hydroxy­2’­deoxyguanosine)
Oxidative stress is known to play an important role in the development of various diseases andaging process. 8­hydroxy­2’­deoxyguanosine (8­OHdG) is formed when DNA is oxidatively
damaged by reactive oxygen species (ROS). 8­OHdG is one of the most sensitive biomarker
for oxidative stress and can be detected in urine, serum, tissue DNA from human and animals.

① High specificity & high sensitivity:
Cross reactivity was checked for 19 analogues.
Formation of 8­hydroxy­2’­deoxyguanosine (8­OHdG) by oxigen radicals.

② Easy operation & speedy:

H Kasai: Environmental Mutagen Research, Vol. 10, p73­78 (1988)

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).

New 8­OHdG Check ELISA (96wells) code: KOG­200SE(Measuring range: 0.5 to 200 ng/mL, suitable for urine sample.)

New 8­OHdG Check Trial Package (32wells) code: KOG­200TE(Measuring range: 0.5 ­ 200 ng/mL, suitable for urine sample.)

Highly Sensitive 8­OHdG Check ELISA (96 wells) code: KOG­HS10E(Measuring range: 0.125 to 10 ng/mL, suitable for serum* and tissue DNA.*)*: sample pretreatment is required.
Reference: S Saito, et.al.: Res. Commun. Mol. Pathol. Pharmacol. 107(1&2), p39­44 (2000)② Anti 8­OHdG monoclonal antibody (clone N45.1)
Highly specific for 8­OHdG.
Anti 8­OHdG mouse monoclonal antibody (IgG1κ), lyophilized powder.

Cross reactivity was checked for 19 analogues. dA, dC, dT, 2’­dG, 8­OHdA, 2’­dI, 2’­dU, O6­methyl­dG, G, 7­methyl­G, 8­Br­G, 8­sulfhydryl­G, 4­SH­G, 8­OH­G, Gua, O6­methyl­Gua, 8­OH­Gua, uric acid,urea, creatinine, creatine (G: Guanosine, Gua: Guanine)

Anti 8­OHdG monoclonal antibody (20 micro gram) code: MOG­020P
Epidermis from hairless mouse byirradiation after 4 weeks of treatment.

chronic

Anti 8­OHdG monoclonal antibody (100 micro gram) code: MOG­100PUVB

Reference: S Toyokuni, et. al.: Lab. Invest. 76(3), p365­374 (1997)③ Anti Thymidine Glycol (TG) monoclonal antibody
ＮＥＷ!

Biomarker for oxidative DNA damage.

Thymidine glycol (TG) is produced when thymidine is damaged by hydroxy radical,
and is a sensitive biomarker for oxidative stress. Anti TG monoclonal antibody specific for TG in
DNA polymers. Suitable for immunohistochemical research.

Anti TG monoclonal antibody (100 micro gram) code: MTG­100P

Oxidative Stress Markers

JaICA④ Anti 4­HNE monoclonal antibody (HNE­J2)→☆

Highly specific for 4­HNE.

→Immunohistochemistry of colorectal carcinoma cells. Adenocarcinoma(arrows→) and nontumorous epithelial cells (star☆;×240). (S. Kondo
et al. Free Radical Biology & Medicine 27 pp401­410, 1999)

Membrane lipids are one of the targets of reactive oxygen species (ROS).4­hydroxy­2­nonenal (4­HNE) is a major membrane lipid peroxidation product. Monoclonal
antibody against 4­HNE (clone HNE­J2) is highly specific for 4­HNE­His/Lys/Cysadducts.
Anti 4­HNE mouse monoclonal antibody (IgG1κ), lyophilized powder.

For immunohistochemistry and western blotting.
OOH

Anti 4­HNE monoclonal antibody (20 micro gram) code: MHN­020P
Anti 4­HNE monoclonal antibody (100 micro gram) code: MHN­100P
Reference: S Toyokuni, et. al.: FEBS Lett. 359, p189­191 (1995)⑤ Hexanoyl­Lysine adduct (HEL) ELISA kit & antibody
A new biomarker of lipid peroxidation.
Hexanoyl­Lysine adduct (HEL) is a novel lipid hydroperoxide­modified lysine residues.
HEL is formed by oxidative modification by oxidized ω­6 fatty acids such as linoleic
acid or arachidonic acid. HEL may be a useful biomarker for initial stage of lipid
peroxidation, and is detected in oxidatively modified LDL, in human atherosclerotic
lesions and human urine.

Lipid hydroperoxide（3­HPODE）

ELISA kit suitable for urine and serum samples.
Nε­(Hexanoyl) Lysine: HEL

Hexanoyl­Lys adduct (HEL) ELISA (96wells) code: KHL­700E(Measuring range: 2 to 700 nmol/L)
Suitable for human urine and animal serum. Required equipments are: pipettes,incubator and microplate reader with 450nm filter.

For immunohistochemistry and western blotting.
Anti HEL monoclonal antibody (20 micro gram) code: MHL­020P
Anti HEL mouse monoclonal antibody (IgG, clone 5H4), lyophilized powder.
Cross reactivity was checked for 8 analogues: MDA, glyoxal, methyl glyoxal,
1­hexanal, 2­hexenal, 1­nonannal, 2­nonenal and 4­hydroxy­2­nonenal.
Reference: Y Kato, et.al.: Biochem. Biophys. Res. Commun. 274, p389­393 (2000)⑥ Anti MDA monoclonal antibody (1F83)
Highly specific for Malondialdehyde (MDA).
Malondialdehyde (MDA) is one of the major aldehyde derived from lipid peroxidation.
This antibody is specific for MDA­modifird proteins, and suitable for
immunohistocheistry and western blotting.

For immunohistochemistry and western blotting.
Anti MDA monoclonal antibody (30 micro gram) code: MMD­030n

JaICA⑦ Test kit for Total Antioxidant Capacity (PAO)
Significance of antioxidants.
For accurate assessment of oxidative stress, measurement of ROS, oxidative
damage and antioxidant activity may be essential. Recently, antioxidants asfunctional foods which scavenge ROS attract a great deal of attention. In the PAO
assay kit, an easy and convenient method to measure antioxidant capacity isprovided. Utilizing the reduction of cupric ion （Cu++⇒Cu+）, antioxidant capacity ofsamples can be detected in 5 minutes.

Application to foods, beverages and serum.
PAO can detect not only hydrophilic antioxidants such as Vitamin C, glutathione, butalso can detect hydrophobic antioxidants such as Vitamin E. Applicable forassessment of total antioxidants of serum, foods and beverage samples.
Sample pretreatment

Samplesrequired

Assay example-１０μL

Mean 1069±145 μmol/L

Human urine

Mix with 3 volumesof distilled water.

２μL

5508 μmol/L

２μL45479 μmol/L

Foods

Mix with 3 volumes Mix with 39of distilled water. volumes ofSample
Mix with 7 volumes
Diluentof distilled water.

２μL

8728 to 46687 μmol/L++

SamplesＣｕReduction by antioxidants+Ｃｕ

Human serum or heparinized plasma

Red wine

Chrolatic reagent(Bathocuproine)

Color 480～490nm(Bathocuproine-Cu+)

Pre-dilution

Green tea
Black tea
CoffeeAssay dilution

Mix with 7 volumesof distilled
Mix withwater.
27
volumes of distilledwater.

２μL
１μL

Potential Antioxidant kit (PAO) (96wells) code: KPA­050E
Suitable for human and animal serum. Required equipments are: pipettes, and microplate
reader with 492nm filter.
Reference: C. VASSALLE, et. al.Journal of Internal Medicine 2004; 256: 308–315⑧ Acrolein (ACR) Antibody
Biomarker of lipid peroxidation.
Acrolein (ACR) is a representative carcinogenc aldehyde found ubiquitously in theenvironment and endogenenously through oxidation reactions, such as lipid
peroxidation and myeloperoxidase­catalyzed amino acid oxidation. ACR is highly
reactive aldehyde and reacts with lysine residue in protein.

For immunohistochemistry and western blotting.
Anti ACR mouse monoclonal antibody (IgG, clone 5F6).
Cross reactivity was checked for 14 analogues: crotonal, 2­hexenal, 2­octanal,2­nonenal, 4­decanal, 2,4­decadional, MDA, 4­HHE, 4­HPE, 4­HON, 4­HNE, n­
propanal, n­pentanal and n­hexenal.

Anti ACR monoclonal antibody (20 micro gram) code: MAR­020n
Anti ACR monoclonal antibody (100 micro gram) code: MAR­100n
Reference: K.Satoh, et. al. Analytical Biochemistry.270,323­328(1999)

Oxidative Stress MarkersJaICA⑨ Anti Dibromo Tyrosine monoclonal antibody (3A5)ＮＥＷ!

A new biomarker of protein oxidation
caused by neutrophils.
Neutrophils and eosinophils play an important role in the defence system against
microbial infection. Myeloperoxidase and eosinophil peroxidase are known tocatalyze formation of hypochlorous acid (HOCl) and hypobromous acid (HOBr).
These reactive intermediates react with proteins, and are known to form tyrosine
halogenation such as dibromotyrosine (DiBrY).
This monoclonal antibody is specific for 3,5­DiBrY, and suitable for
immunohistochemical analysys of oxidative stress.

Anti DiBrY monoclonal antibody (20 micro gram) code: MBY­020P
Reference: Y Kato, et. ak. Rad Biol Med, 38, p24­31 (2005)⑩ Other antibodies for Lipid peroxidation markers
Anti 4­HHE monoclonal antibody

Anti MG monoclonal antibody(4­hydroxy­2­hexenal)(Methylglyoxal)

4­hydroxy­2­alkenal is one of the major lipid peroxidation products,and shows many biological effects such as high toxicity to cells.
Among them, 4­hydroxy­2­hexenal (4­HHE) is an aldehyde formed
during peroxidation of n­3 fatty acids such as docosahexaenoicacid.

Methylglyoxal (MG), an endogenous metabolite that increases indiabetes and is a common intermediate in the Maillard reaction(glycation), reacts with proteins and forms advanced glycation end
products (AGE).

Anti 4­HHE monoclonal antibody (30 micro gram)Anti MG monoclonal antibody (30 micro gram)code: MHH­030n

code: MMG­030n

Anti 7­KC monoclonal antibody

Anti CRA monoclonal antibody(7­ketocholesterol)(Crotonaldehyde)

Cholesterol oxidation products, especially 7­ketocholesterol (7­KC)
have been the focus of much attention because they are present
in human atheroscleotic plaque and display a wide range ofatherogenic properties in vitro and in vivo.

Crotonaldehyde (CRA) is a representative carcinogenic aldehyde
formed endogenously through lipid peroxidation. CRA is a highly
reactive aldehyde and reacts with lysine residue in proteins.

Anti 7­KC monoclonal antibody (20 micro gram)Anti CRA monoclonal antibody (30 micro gram)code: MKC­020n

code: MCA­030n

Developed by & Technical support:

Japan Institute for the Control of Aging (JaICA),
Nikken SEIL Co., Ltd.

Haruoka 710­1, Fukuroi, Shizuoka, 437­0122 Japanbiotech@jaica.com Phone: +81­538­49­0125 FAX: +81­538­49­1267

http://www.jaica.com/

Established in 1985. A satellite center of Trace Element Institute for UNESCO.
These products are developed and manufactured in Japan. For research use only. ©2005­2008 JaICA