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To general oxidative stress markers in the body.

References and Technical Information

8-OHdG top

New/Highly Sensitive 8-OHdG Check Applicaion to urine, serum and tissue samples.
Anti 8-OHdG monoclonal antibody Application to immunohistochemistry.

No. Title Content
1 Animals applicable Q: What kind of animals is applicable to 8-OHdG ELISA?
A: Applicable to urine samples from human, mouse, rat, rabbit, dog, cattle, horse, etc. Please remove cells and insluble materials from urine before application to ELSIA. To obtain more reliable data, we recommend to dilute urine samples with 1 to 10 volumes of phosphate buffered saline (PBS pH7.4).
For urine samples, "New 8-OHdG Check" is suitable. For serum and tissue samples, we recommend to apply to "Highly Sensitive 8-OHdG Check" after pretreatment of samples.
2 Sample dilution Q: What should be used for sample dilution?
A: Phoshate buffered saline (PBS, pH7.4) is recommended. If urine sample is from healthy human, sample can be applied to "New 8-OHdG Check ELISA" as it is, but if urine sample is from patients, we recommend to dilute samples for 2 or 4 times with PBS.
3 Stability of urinary 8-OHdG Q: How urine samples should be stored?
A: 8-OHdG is known to chemically stable. Urine sample can be stored for 3 days at room temperature, for 7 days at 4 degree C. But please take care for contamination of bacteria. If hydrochloric acid is used to prevent bacterial growth, please adjust pH before apply to ELISA.
Urine samples can be stored for longer period, for years at -80 degree C. But please avoid repeated freeze / thaw cycle. When frozen urine sample is thaw, insolbule materials may be observed in some cases. Please remove insoluble materials by centrifuge before application to ELISA.
4 Type of samples applicable Q: What kind of samples are applied to 8-OHdG ELISA?
A: In addition to urine samples, serum / plasma, ascite samples, outer liquid of artificial dialysis and saliva samples are known to be applied to 8-OHdG ELISA. Pretreatment is required for these samples. DNA samples isolated from blood cells, sperm, organs and cultured cells can be applied to 8-OHdG ELISA.

[Reference]
New biomarker evidence of oxidative DNA damage in whole saliva from clinically healthy and periodontally diseased individuals. M Takane, et. al., J Periodontol.73(5), p551-554 (2002)
5 Mean concentration of urinary 8-OHdG
(Normal human)
A) Mean concentration: 13.6 ng/mL (250 males) and 10.3 ng/mL (250 females)
B) 8-OHdG extretion ratio: 8.6 ng/kg/h (250 males and 250 females)
C) Creatinin correction: 8.4 ng/mg CRE (250 males and 250 females)
6 Urine sampling and correction methods 8-OHdG concentraion in random urine is not stable, depending on individual variation, the time in the day of sample collection, sex and some other factors. To obtain the best result, lease take care for sampling and correction methods.

About creatinine correction:
Creatinie correction is known to be a good method to obtain stable data. But it is not suitable for some researchers who are interested in the effect of exercise to oxidative stress. That's because creatinine concentration will be increased by exercise.

1) Effect of repeated exercise on urinary 8-hydroxy-deoxyguanosine excretion in humans. Free Radic Res. 1997 Jun;26(6):507-14. Okamura K, et. al.
In this paper, effects of running are assessed using 8-OHdG excretion per day. A significant increase of urinary 8-OHdG is observed (from 265.7+/-75.5 to 335.6+/-107.4 pmol/ kg/ day).

2) Habitual long-distance running does not enhance urinary excretion of 8-hydroxydeoxyguanosine. Eur J Appl Physiol Occup Physiol. 1997;75(5):467-9. Pilger A,et. al.
In this paper, effects of running are assessed by urinary 8-OHdG with creatinine correction. Both 8-OHdG and creatinine concentration increased. But the effect of running to 8-OHdG level is not significant (0.12 - 6.45 umol/mol CRE).
7 Serum / plasma samples: To detect 8-OHdG in serum and plasma samples, pretreatment is required. Please remove proteins by ultrafiltration, and apply to "Highly Sensitive 8-OHdG Check".
8 Blood collection tubes: Blood collection tubes are commercially available. For example, NIPRO code.30-122 is suitable. Please note that serum 8-OHdG concentration may be increased if whole blood is placed for hours at room temperature. We recommend to start centrifuge in 20 minutes after blood collection. Serum samples must be stored below -20 degree C.
9 Information about human serum 8-OHdG level:
8-OHdG concentration in male serum is higher than that of female. Serum 8-OHdG is higher in smokers than in non-somokers.
Reference:The relationship between smoking habits and serum levels of 8-OHdG, oxidized LDL antibodies, Mn-SOD and carotenoids in rural Japanese residents., J Epidemiol 2003 Jan;13(1):29-37.Suzuki K, et.al.
List of references
1 T.Osawa, A.Yoshida, S.Kawakishi, Y.Yamashita, and H.Ochi:
Protective role of dietary anti-oxidants in oxidative stress.
Oxidative Stress and Aging (Esd:R.G.Cutler, L.Packer, Bertram and A.Mori) p367-377, Birkhuser Verlag Basel, Switzerland (1995)
2 J.Leinonen, T.Lehtimki, S.Toyokuni, K.Okada, T.Tanaka, H.Hiai, H.Ochi, P.Laippala, V.Rantalaiho, O.Wirta, A.Pasternack, and H.Alho:
New biomarker evidence of oxidative DNA damage in patients with non-insulin-dependent diabetes mellitus.
FEBS Lett. 417, p150-152 (1997)
3 M.Erhola, S.Toyokuni, K.Okada, T.Tanaka, H.Hiai, H.Ochi, K.Uchida, T.Osawa, M.M.Nieminen, H.Alho, and P.K-Lehtinen:
Biomarker evidence of DNA oxidation in lung cancer patients: association of urinary 8-hydroxy-2?Edeoxyguanosine excretion with radiotherapy, chemotherapy, and response to treatment.
FEBS Lett. 409, p287-291 (1997)
4 H.Tsuboi, K.Kouda, H.Takeuchi, M.Takigawa, Y.Masamoto, M.Takeuchi, and H.Ochi:
8-Hydroxydeoxyguanosine in urine as an index of oxidative damage to DNA in the evaluation of atopic dermatitis.
Br. J. Dermatol. 138, p1033-1035 (1998)
5 Y.Miyake, K.Yamamoto, N.Tsujihara, and T.Osawa:
Protective effects of lemon flavonoids on oxidative stress in diabetic rats.
Lipids 33(7), p689-695 (1998)
6 M-H.Kang, M.Naito, N.Tsujihara, and T.Osawa:
Sesamolin inhibits lipid peroxidation in rat liver and kidney.
J. Nutr. 128, p1018-1022 (1998)
7 S.S.Kantha, S.Wada, H.Tanaka, M.Takeuchi, S.Watabe, and H.Ochi:
Carnosine sustains the retention of cell morphology in continuous fibroblast culture subjected to nutritional insult.
Biochem. Biophys. Res. Commun. 223, p278-282 (1996)
8 S.S.Kantha, S.Wada, M.Takeuchi, S.Watabe, and H.Ochi:
A sensitive method to screen for hydroxyl radical scavenging activity in natural food extracts using competitive inhibition ELISA for 8-hydroxydeoxyguanosine.
Biotechnol. Techniques 10(12), p933-936 (1996)
9 T.Arimoto, T.Yoshikawa, H.Takano and M.Kohno:
Generation of rective oxygen species and 8-hydroxy-2'-deoxyguanosine formation from diesel exhaust particles components in L1210 cells.
Japanese J. Pharmacol. 80, p49-54 (1999)
10 M.D.Evans, M.S.Cooke, I.D.Podmore, Q.Zheng, K.E.Herbert, and J.Lunec:
Discrepancies in the measurement of UVC-induced 8-oxo-2'-deoxyguanosine: Implications for the analysis of oxidative DNA damage.
Biochem. Biophys. Res. Commun. 259, p374-378 (1999)
11 M.S.Cooke, M.D.Evans, I.D.Podmore, K.E.Herbert, N.Mistry, P.Mistry, P.T.Hickenbotham, A.Hussieni, H.R.Griffiths, and J.Lunec:
Novel repair action of vitamin C upon in vivo oxidative DNA damage.
FEBS Lett. 363, p363-367 (1998)
12 M.S.Cooke, M.D.Evans, K.E.Herbert, and J.Lunec:
Urinary 8-oxo-2'-deoxyguanosine - source, significance and supplements.
Free Rad. Res. 32, p381-397 (2000)
13 T.Yoshikawa, L.L-Fen, Y.Naito, N.Yoshida, T.Osawa, H.Ochi and M.Kondo:
Rapid detection of 8-hydroxy-2?Edeoxyguanosine formed by hydrogen peroxide in cultured cells by using an enzyme-linked immunosorbent assay.
(In press)
14 T. Shimoike, T. Inoguchi, F. Umeda, H. Nawata, K. Kawano and H. Ochi:
The meaning of serum levels of advanced glycosylation end products in diabetic nephropathy.
Metabolism 49(8), p1030-1035 (2000)
15 W. Y. Fan, K. Ogusu, K. Kouda, H. Nakamura, T. Satoh, H. Ochi and H. Takeuchi:
Reduced oxidative DNA damage by vegetable juice intake: A controlled trial.
J. Physiol. Anthropol. 19(6), p287-289 (2000)
16 H.Ochi, M.Hashimoto, J.Kurashige:
Assessment of functional tea in human using oxidative stress profile technique.
Proceedings of 2001 International Conference on O-Cha (tea) Culture and Science (2001)
17 T. Kuragano, T. Kuno, C. Yamamoto, Y. Nagura, S. Takahashi and K. Kanmatsuse:
Oxidative stress on DNA in chronic renal failure: the influence of different hemodialysis membranes.
J. Artif. Organs 4, p320-325 (2001)
18 M.S.Cooke, M.D.Evans, R.M.Burd, K.Patel, A.Barnard, J.Lunec and P.E.Hutchinson:
Induction and excretion of ultraviolet-induced 8-oxo-2'deoxyguanosine and thymine dimmers in vivo: implications for PUVA.
J. Invest. Dermatol. 116(2), p281-285 (2001)
19 M.S.Cooke, M.D.Evans and J.Lunec:
DNA repair: insights from urinary lesion analysis.
Free Rad. Res. 36(9), p929-932 (2002)
20 M.S.Cooke, M.D.Evans, N.Mistry and J.Lunec:
Role of dietary antioxidants in the prevention of in vivo oxidative DNA damage.
Nutr. Res. Rev. 15, p19-41 (2002)
21 T. Matsubasa, T. Uchino, S. Karashima, Y. Kondo, K. Maruyama, M. Tanimura and F. Endo:
Oxidative stress in very low birth weight infants as measured by urinary 8-OHdG.
Free Rad. Res. 36(2), p189-193 (2002)
22 Y.Ibuki, T.Warashina, T.Noro and R.Goto:
Coexposure to benzo[a]pyrene plus ultraviolet A induces 8-oxo-7,8-dihydro-2?Edeoxyguanosine formation in human skin fibroblasts: preventive effects of anti-oxidant agents.
Environ. Toxicol. Pharmacol. 12, p37-42 (2002)
23 M.Kakimoto, T.Inoguchi, T.Sonta, H.Y.Yu, M.Imamura, T.Etoh, T.Hashimoto and H.Nawata:
Accumulation of 8-hydroxy-2?Edeoxyguanosine and mitochondrial DNA deletion in kidney of diabetic rats.
Diabetes 51, p1588-1595 (2002)
24 Takane, Naoyuki Sugano, Hiroyasu Iwasaki, Yoshihiro Iwano, Noritaka Shimizu and Koichi Ito:
New biomarker evidence of oxidative DNA damage in whole saliva from clinically healthy and periodontally diseased individuals.
J.Periodontol 73(5), p551-554 (2002)
25 Kayoko Shimoi, Hiroshi Kasai, Naoko Yokota, Shinya Toyokuni and Naohide Kinae:
Comparison between high-performance liquid chromatography and enzyme-linked immunosorbent assay for the determination of 8-hydroxy-2?Edeoxyguanosine in human urine.
Cancer Epidemiol. Biomarkers & Prevention 11 p767-770 (2002)
26 Ha Won Kim, Akira Murakami, Marshall V Williams and Hajime Ohigashi:
Mutagenicity of reactive oxygen and nitrogen species as detected by co-culture of activated inflammatory leukocytes and AS52 cells.
Carcinogenesis 24(2), p235-241 (2003)
27 Nagayoshi Y, Kawano H, Hokamaki J, Miyamoto S, Kojima S, Shimomura H, Tsujita K, Sakamoto T, Yoshimura M, Ogawa H:
Urinary 8-hydroxy-2'-deoxyguanosine levels increase after reperfusion in acute myocardial infarction and may predict subsequent cardiac events.
Am J Cardiol. 95(4), p514-517 (2005)
28 Yutaka Otsuji, Eiji Kuwahara, Keiko Yuge, Goichi Yotsumoto, Takayuki Ueno, Kenichi Nakashiki, Shuichi Hamasaki, Sadatoshi Biro, Shinichi Minagoe, Robert A. Levine, Ryuzo Sakata, and Chuwa Tei:
Relation of Aneurysmectomy in Patients With Advanced Left Ventricular Remodeling to Postoperative Left Ventricular Filling Pressure, Redilatation With Ischemic Mitral Regurgitation.
Am J Cardiol 95, p517-521 (2005)

List of references
1 T.Tanaka, Y.Nishiyama, K.Okada, K.Hirota, M.Matsui, J.Yodoi, H.Hiai, and S.Toyokuni:
Induction and nuclear translocation of thioredoxin by oxidative damage in the mouse kidney: independence of tubular necrosis and sulfhydryl depletion.
Lab. Invest. 77(2), p145-155 (1997)
2 Y.Hattori, C.Nishigori, T.Tanaka, K.Uchida, O.Nikaido, T.Osawa, H.Hiai, S.Imamura, and S.Toyokuni:
8-Hydroxy-2'deoxyguanosine is increased in epidermal cells of hairless mice after chronic ultraviolet B exposure.
J. Invest. Dermatol. 107, p733-737 (1997)
3 S.Toyokuni, T.Tanaka, Y.Hattori, Y,Nishiyama, A.Yoshida, K.Uchida, H.Hiai, H.Ochi, and T.Osawa:
Quantitative immunohistochemical determination of 8-hydroxy-2'deoxyguanosine by a monoclonal antibody N45.1: Its application to ferric nitrilotriacetate-induced renal carcinogenesis model.
Lab. Invest. 76(3), p365-374 (1997)
4 S.Takahashi, M.Hirose, S.Tamano, M.Ozaki, S.Orita, M.Takeuchi, H.Ochi, S.Fukuda, H.Kasai, and T.Shirai:
Immunohistochemical detection of 8-hydroxy-2?Edeoxyguanosine in paraffin-embedded sections of rat liver after carbon tetrachloride treatment.
Toxicol. Pathol. 26, p247-252 (1998)
5 N.U.Ahmed, M.Ueda, O.Nikaido, T.Osawa and M.Ichihashi:
High levels of 8-hydroxy-2'deoxyguanosine appear in normal human epidermis after a singer dose of ultraviolet radiation.
Br. J. Dermatol. 140, p226-231 (1999)
6 Y.Ihara, S.Toyokuni, K.Uchida, H.Odaka, T.Tanaka, H.Ikeda, H.Hiai, Y.Seino, and Y.Yamada:
Hyperglycemia causes oxidative stress in pancreatic -cells of GK rats, a model of type 2 diabetes.
Diabetes 48, p927-932 (1999)
7 S.Kondo, S.Toyokuni, Y.Iwasa, T.Tanaka, H.Onodera, H.Hiai and M.Imamura:
Persistent oxidative stress in human colorectal carcinoma, but not in adenoma.
Free Rad. Biol. Med. 27, p401-410 (1999)
8 S.Toyokuni, Y.Iwasa, S.Kondo, T.Tanaka, H.Ochi, and H.Hiai:
Intranuclear distribution of 8-hydroxy-2'-deoxyguanosine: An immunocytochemical study.
J. Histochem. Cytochem. 47(6), p833-835 (1999)
9 S.Toyokuni:
Reactive oxygen species-induced molecular damage and its application in pathology.
Pathol. Internat. 49, p91-102 (1999)
10 S.Toyokuni:
An in vitro Feton reaction model and the application of immunohistochemistry to detect oxidative damage.
Models and Methods in Cell Signaling and Gene Expression: Applications to Oxidative Stress Research., Chapter four pp40-54 (2000)
11 K.Yamagami, Y.Yamamoto, M.Kume, Y.Ishikawa, Y.Yamaoka, H.Hiai and S.Toyokuni:
Formation of 8-hydroxy-2'deoxyguanosine and 4-hydroxy-2-nonenal-modified proteins in rat liver after ischemia-reperfusion: Distinct localization of the two oxidatively modified products. Antioxidants & Redox Signaling, 2(1), p127-136 (2000)
12 D.Nakae, H.Akai, H.Kishida, O.Kusuoka, M.Tsutsumi, and Y.Konishi:
Age and organ dependent spontaneous generation of nuclear 8-hydroxydeoxyguanosine in male Fischer 344 rats.
Lab. Invest. 80(2), p249-261 (2000)
13 S.Liardet, C.Scaletta, R.Panizzon, P.Hohlfeld and L.L.Applegate:
Protection against pyrimidine dimmers, p53, and 8-hydroxy-2'deoxyguanosine expression in ultraviolet-irradiated human skin by sunscreens: difference between UVB + UVA and UVB alone sunscreens.
J. Investigative Dermatol. 117(6), p1437-1441 (2001)