Japan Institute for the Control of Aging: JaICA Oxidative Stress
Rev.081027
Anti-8-OHdG Monoclonal Antibody (clone N45.1)
Anti- 8-hydroxy-2'-deoxyguanosine monoclonal antibody
FOR RESEARCH USE ONLY
About 8-hydroxy-2'-deoxyguanosine (8-OHdG):
Reactive oxygen species (ROS) and oxidative stress may be crucial for development of various diseases such as diabetes, cancer, and may play an important role in the aging process. 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. Anti 8-OHdG monoclonal antibody (clone N45.1) is highly specific for DNA damage, not cross react with RNA oxidation products such as 8-hydroxy-guanine and 8-hydroxy-guanosine. Suitable for immunohistochemistry and ELISA.
Epidermis from hairless mouse by chronic UVB irradiation after 4 weeks of treatment stained with N45.1.
(Y.Hattori, et.al.: J Invest Dermatol 107, p733-737, 1996)
Specifications
Clone #: N45.1
Antigen: 8-OHdG-conjugated Keyhole Limpet Hemocyanin
Subclass: Mouse IgG1(kappa)
Prepared as ascite, and ammnonium sulphate purified.
Form: Lyophilized Powder
Package: 20 or 100 µg of IgG / vial (MOG-020P and MOG-100P respectively).
Specificity: 19 analogues of 8-OHdG {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-OHGua, uric acid, Urea, creatine, creatinine} demonstrate no cross-reactivity. Only 8-sulfhydryl-G and 8-OHG demonstrate minimal cross-reactivity (less than 1%).
Use: Immunohistochemistry and ELISA
Product name Code Content
Anti 8-OHdG monoclonal antibody MOG-020P 20 µg of IgG
MOG-100P 100 µg of IgG
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
 
References
1) 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.]
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 (1996)
[ Immunohistochemistry of UV-B irradiated mouse skin. ]
3) 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)
[ Immunohistochemistry of ferric nitrotriacetate-treated mouse kidney. ]
4) 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)
[ Immunohistochemistry of pancreatic islets from GK rat, a diabetes model. ]
5) T. Ichiseki, A. Kaneuji, S. Katsuda1, Y. Ueda,T. Sugimori and T. Matsumoto: DNA oxidation injury in bone early after steroid administration is involved in the pathogenesis of steroid-induced osteonecrosis. Rheumatology 44,p456-460(2005)
[ Immunohistochemistry of rabbit tissue. ]
6) Lee YA, Cho EJ, Yokozawa T: Protective effect of persimmon (Diospyros kaki) peel proanthocyanidin against oxidative damage under H2O2 -induced cellular senescence. Biol Pharm Bull 31(6)p1265-1269(2008)
[ Application to human cultured cells. ]
Immunohistochemical procedure
Fixation: For formalin-fixed tissue and cultured cells, antigen retrieval process may be useful.
Tissue samples are fixed by Bouins Solution, antigen retrieval process will not be needed.
Blocking: Apply normal rabbit serum (Diluted to 1:75) (DAKO corporation, Kyoto, Japan) to the specimens
for the inhibition of non-specific binding of secondary antibody.
If sample tissues are from mouse, blocking of internal mouse IgG will be needed.
Primary antibody: Apply N45.1 (1-10microgram/ml) to the specimens, incubate overnight at 4 degree C.
Secondary antibody: Apply biotin-labeled rabbit anti-mouse IgG serum (Dako; Diluted to 1:300) for 40min at room temperature.
ABC method: Apply avidin-biotin-alkaline phosphatase complex (Vector Lab., Burlingame, CA; Diluted to 1:100)
for 40min at room temperature.
Staining: Use a kit of black substrate for alkaline phosphatase (BCIP/NBT, Vector).
  Antigen retrieval:
Put sample slides into 500L glass beaker with 10mM citrate buffer pH6.0 and heat.
Keep boiling for 5 minutes.
Cool down slowly in 1 hour.
 
 
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.