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. New 8-OHdG Check is a competitive enzyme-linked immunosorbent assay (ELISA) utilising monoclonal antibody (clone N45.1) which is highly specific for DNA damage, 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.

This product is a 8-OHdG ELISA kit utilizing which is highly specific for 8-OHdG. We provide two types of 8-OHdG ELISA kits with different assay range. New 8-OHdG Check ELISA is suitable for urine and serum sample from animal and human.

Assay principle:

Competitive ELISA (detection: 450 nm)

Specifity:

Specific for 8-OHdG. Antibody have been tested to 8-OHdG analogues (guanosine(G),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-OH-Gua, uric acid, urea, creatine, creatinine, 8-sulfhydryl-G, 8-OH-G).

Measuring range:

0.5 to 200 ng/mL

Format:

96 wells (18 samples in triple assays)

Applications:

Urine from human and animals. 
 

Storage:

Store at 4 - 10°C (don't freeze).

Expiry:

12 months after the day of manufacturing.

Required but not provided:

Micropipet and chip (100 micro L, 1000 micro L).
Measuring pipet (10 mL, 20 mL)/ measuring cylinders.
8 or 12-syncronous multichannel pipet and reagent tray for multichannel pipet.
Microplate reader (filter; 450 nm).

8-OHdG Microtiter Plate:

Precoated with 8-OHdG(12 X 8wells, split type)^*

1 plate

Primary Antibody:

Anti 8-OHdG antibody, powder.

1 vial

Primary Antibody Solution

1 vial (6mL)

Secondary Antibody:

HRP-anti mouse antibody, powder.

1 vial

Secondary Antibody Solution:

1 vial (12mL)

Chromatic Solution:

3,3',5,5'-tetramethylbenzidine

1 vial (0.25mL)

Diluting Solution:

H₂O₂ containing buffer.

1 vial (12mL)

Washing Solution(5x):

2 vials (26mL x 2)^*

Reaction Terminating Solution:

1M Phosphoric acid.

1 vial (12mL)

Standard 8-OHdG Solution:

Purified 8-OHdG (0.5, 2, 8, 20, 80, 200 ng/mL)

1 vial each

Plate Seal:

2 sheets

*:Trial package (KOG-200TE) contains 32 wells (4 X 8 wells), and one vial of Washing solution(5X).

1)

H.Kasai, P.F.Crain, Y.Kuchino, S.Nishimura,A.Ootsuyama and H.Tanooka : Formation of 8-hydroxyguaine moiety in cellular DNA by agents producing oxygen radicals and evidence for its repair. Carcinogenesis 7(11), p1849-1851 (1986)
[ 8-OHdG/8-OHG is formed by reactive oxygen radicals. ]

2)

R.G.Cutler: Antioxidants and aging. Am J Clin Nutr 50,p373S-379S (1991)
[ Relationship between lifespan and antioxidants, enzymes and 8-OHdG. ]

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)
[ Specifity of the antibody clone N45.1.]

4)

Saito S, Yamauchi H, Hasui Y, Kurashige J, Ochi H, Yoshida K: Quantitative determination of urinary 8-hydroxydeoxyguanosine (8-OH-dg) by using ELISA. Res Commun Mol Pathol Pharmacol 107(1-2),p39-44 (2000)
[ Development and assessment of 8-OHdG Check ELISA. ]

5)

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)
[ Measurement of 8-OHdG in human urine. ]

6)

Morillas-Ruiz J, Zafrilla P, Almar M, Cuevas MJ, Lopez FJ, Abellan P, Villegas JA, Gonzalez-Gallego J: The effects of an antioxidant-supplemented beverage on exercise-induced oxidative stress: results from a placebo-controlled double-blind study in cyclists. Eur J Appl Physiol 95(5-6),p543-549(2005)
[ Study about exercise, antioxidant supplements and urinary 8-OHdG. ]

7)

Someya T, Kaneko K, Yamada T, Yamashiro Y: Effect of a novel free radical scavenger, edaravone, on puromycin aminonucleoside induced nephrosis in rats. Pediatr Nephrol 20(10),p1430-1434(2005)
[ Effect of antioxidants to 8-OHdG in rat urine. ]

8)

Shiihara T, Kato M, Ichiyama T, Takahashi Y, Tanuma N, Miyata R, Hayasaka K: Acute encephalopathy with refractory status epilepticus: Bilateral mesial temporal and claustral lesions, associated with a peripheral marker of oxidative DNA damage. J Neurol Sci 250(1-2),p159-161(2006)
[ Measurement of 8-OHdG in human urine, serum and cerebro spinal fluid(CSF). ]