Genox

Anti-8-Hydroxy-2’-Deoxy Guanosine
Monoclonal Antibody

(Catalog #MOG-100 for 100 microgram vial)
(Catalog #MOG-020 for 20 microgram vial)
(Anti 8-OHdG monoclonal antibody, clone N45.1)

Product Insert (pdf)pdf

What  is 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 8-OHdG, and
is suitable for immunohistochemistry.


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, 1997)

 

 

Clone #

:

N45.1

Antigen

:

8-OHdG-conjugated Keyhole Limpet Hemocyanin

Subclass

:

Mouse IgG1(kappa)
Prepared as ascite, and ammnonium sulphate purified.

Form

:

Lyophilized Powder

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

Storage

:

Less than -20°C for 5 years
When transport, stable at least 7 days at room temperature.


IMMUNOHISTOCHEMISTRY

This is an application example for anti 8-OHdG monoclonal antibody (clone N45.1). Experimental conditions should be determined individually by each user.

      
1) Fixation:

Fix the fresh tissues with Bouin’s Solution (Saturated Picric Acid: Formaldehyde: Acetic Acid : H2O = 15:5:1:10) overnight.
Please note that use of specimens fixed in the same conditions are recommended for quantitative analyses.

2)  Deparaffinization (In case of paraffin embedded specimens):

Deparaffin with xylene and ethanol.

3)  Antigen Retrieval (In case of fixation with formaldehyde): 

Microwave method or Autoclave method is recommended.

4) Avidin Biotin Complex (ABC) Method:

a)
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.
b)
Apply N45.1 (1-10microgram/ml) to the specimens, incubate overnight at 4 degree C.
c)
Apply biotin-labeled rabbit anti-mouse IgG serum
(Dako; Diluted to 1:300) for 40min at room temperature.
d)
Apply avidin-biotin-alkaline phosphatase complex (Vector Lab., Burlingame, CA; Diluted to 1:100) for 40min at room temperature.
e)
Staining. Use a kit of black substrate for alkaline phosphatase (Vector).

 

 

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'-deoxy-guanosine by a monoclonal antibody N45.1: Its application to ferric nitrilotriacetate-induced renal carcinogenesis model.
Laboratory Investigation, 76(3), 365-374 (1997)

2)

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)

3)

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)

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’-deoxyguanosine in paraffin-embedded sections of rat liver after carbon tetrachloride treatment.
Toxicol. Pathol. 26, p247-252 (1998)

5)

S.Toyokuni.  Reactive oxygen species-induced molecular damage and its application in pathology.
Pathology International, 49, 91-102 (1999) 

6)

S.Toyokuni, Y.Iwasa, S.Kondo, T.Tanaka, H.Ochi, and H.Hiai.  Intranuclear distribution of 8-hydroxy-2'-deoxyguanosine: An immunocytochemical study.
The Journal of Histochemistry and Cytochemistry, 47(6), 833-835 (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 Radical Biology and Medicine, 21(3/4), 401-410 (1999)  

8)

Y.Ihara, S.Toyokuni, K.Uchida, H.Okada, 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, 927-932 (1999)

9)

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)

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)

 

The ‘Anti-8-OHdG Monoclonal Antibody’ is manufactured in Japan
by the Japan Institute for the Control of Aging (JalCA). Genox is the distributor of this product for users in North and South America.

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