Highly Sensitive 8-OHdG Check ELISA

8-oxo-dG/8-hydroxy-2'-deoxyguanosine

Suitable for assessment of oxidative stress in serum, tissue and cultured cells. For research use only.
About 8-hydroxy-2'-deoxyguanosine (8-OHdG):

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 biomaterialsfrom human and animals.
This product is a 8-OHdG ELISA kit utilizing anti 8-OHdG monoclonal antibody (clone N45.1) which is highly specific for 8-OHdG. We provide two types of 8-OHdG ELISA kits with different assay range. Highly Sensitive 8-OHdG Check ELISA is suitable for urine, serum , tissue and cultured cells.

Specifications

1. Assay principle: Competitive ELISA (detection: 450 nm)
2. 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).
3. Measuring range: 0.125 to 10 ng/mL
4. Format: 96 wells (18 samples in triple assays)
5. Applications: Urine, serum, tissue and cultured cells.
6. Storage: Store at 4 - 10°C (don't freeze).
7. Expiry: 9 months after the day of manufacturing.
8. Required but not provided: Micropipet and chip (100 µL, 1000 µL).
9. Measuring pipet (10 mL, 20 mL)/ measuring cylinders.
10. 8 or 12-syncronous multichannel pipet and reagent tray for multichannel pipet.
11. Microplate reader (filter; 450 nm).

Content of this kit

1. 8-OHdG Microtiter Plate: Precoated with 8-OHdG(12 X 8wells, split type) 1 plate
2. Primary Antibody: Anti 8-OHdG antibody, powder. 1 vial
3. Primary Antibody Solution 1 vial (6mL)
4. Secondary Antibody: HRP-anti mouse antibody, powder. 1 vial
5. Secondary Antibody Solution: 1 vial (12mL)
6. Chromatic Solution: 3,3',5,5'-tetramethylbenzidine 1 vial (0.25mL)
7. Diluting Solution: H2O2 containing buffer. 1 vial (12mL)
8. Washing Solution(5x): 2 vials (26mLx2)
9. Reaction Terminating Solution: 1M Phosphoric acid. 1 vial (12mL)
10. Standard 8-OHdG Solution: Purified 8-OHdG (0.125, 0.25, 0.5, 1, 4, 10 ng/mL) 1 vial each
11. Plate Seal: 2 sheets

References

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) 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)
[ ELISA results show high correlation between those of HPLC-ECD when 8-OHdG in DNA samples are assessed. ]
6) 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)
[ Detection of 8-OHdG in cultured cells. ]
7) Watanabe E, Matsuda N, Shiga T, Kajimoto K, Ajiro Y, Kawarai H, Kasanuki H, Kawana M: Significance of 8-hydroxy-2'-deoxyguanosine levels in patients with idiopathic dilated cardiomyopathy. J Card Fail. 2006 Sep;12(7):527-32.
[ Serum 8-OHdG in IDC patients are assessed. ]
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):159-61(2006)
[ Measurement of 8-OHdG in human urine, serum and cerebro spinal fluid(CSF). ]
9) Reiko Nagasaka, Nobuaki Okamoto, Hideki Ushio: Effects of caloric restriction on post-spawning death of ayu. Exp Gerontol 40, p556-561(2005)
[ Detection of 8-OHdG in fish tissues (brain and liver). ]
10) Yasuda M, Ide H, Furuya K, Yoshii T, Nishio K, Saito K, Isotani S, Kamiyama Y, Muto S, Horie S: Salivary 8-OHdG: a useful biomarker for predicting severe ED and hypogonadism. J Sex Med. 5(6),p1482-1491(2008)
[ 8-OHdG in saliva can be detected by 8-OHdG ELISA. ]

Product name Code Assay range Application
Highly Sensitive 8-OHdG Check KOG-HS10E 0.125 - 10 ng/mL Serum, plasma, tissue and other biological fluids.

Made in Japan.