Genox: Anti-4-Hydroxy-2-Nonenal Monoclonal Antibody

Anti-4-Hydroxy-2-Nonenal Monoclonal Antibody
(Anti 4-HNE monoclonal antibody, clone HNEJ-2)
(Catalog #MHN-100 for 100 microgram vial)
(Catalog #MHN-020 for 20 microgram vial)

Product Insert (pdf)

What is 4-HNE?
Reactive oxygen species (ROS) are involved in lipid peroxidation inside the living bodies, and membrane lipids are one of the major targets of ROS. During the peroxidation process, a variery of aldehydes are formed. 4-hydroxy-2-nonenal (4-HNE) is an alpha, beta unsaturated aldehyde that can be formed by peroxidation of omega-6 unsaturated fatty acids such as linoleic acid and arachidonic acid. Especially, 4-HNE formed inside living body is reported to originate from phospholipid-bound arachidonic acid. 4-HNE may be one of the most reliable biomarker of lipid peroxidation.

Structure of 4-Hydroxy-2-Nonenal

Immunohistochemcal detection of 4-HNE in colorectal carcinoma cells.
Adenocarcinoma (arrows) and non tumorous epithelial cells (star).
(S. Kondo, et al.:Free Radical Biology & Medicine 27, p401-410, 1999)

Clone # Immunogen	:	HNEJ-2 4-HNE-modified Keyhole Limpet Hemocyanin
Subclass	:	Mouse IgG1(kappa) Prepared as ascite, and ammnonium sulphate purified.
Form	:	Lyophilized Powder (containing 20ug or 100ug of IgG)
Specificity	:	This antibody show almost negligible reactivity with proteins that were treated with other aldehydes, such as 2-nonenal, 2-hexenal, 1-hexanal, 4-hydroxy-2-hexenal,formaldehyde, or glutaraldehyde.
Applications	:	Immunohistochemistry and western blotting.
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 4-hydroxy-2-nonenal (4-HNE) monoclonal antibody, clone HNE-J2. (Catalog #MHN-020P/ and MHN-100P).

1) Fixation:

Fixation using Bouin's solution is recommended.
In this case, antigen retrieval process is not required.

2) 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).

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

Microwave method or Autoclave method is recommended.

REFERENCES

1)	S.Toyokuni, N.Miyake, H.Hiai, M.Hagiwara, S.Kawakishi, T.Osawa, and K.Uchida The monoclonal antibody specific for the 4-hydroxy-2-nonenal histidine adduct. FEBS Letters, 359, 189-191 (1995)
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. The Journal of Investigative Dermatology, 107, 733-737 (1997)
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. Laboratory Investigation, 77(2), 145-155 (1997)
4)	S.Toyokuni. Reactive oxygen species-induced molecular damage and its application in pathology. Pathology International, 49, 91-102 (1999)
5)	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)
6)	T.D.Oberley, S.Toyokuni, and L.I.Szweda. Localization of hydroxynonenal protein adducts in normal human kidney and selected human kidney cancers. Free Radical Biology and Medicine, 27(5/6), 695-703 (1999)

The ‘Anti-4-Hydroxy-2-Nonenal 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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Note: Genox’s Products are for RESEARCH USE ONLY.
Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS.

The provision of test reports, generated by using Genox's products, to individuals
or to treating physicians for the diagnosis, prevention, treatment and control of any human disease or impairment of, or the assessment of the health, nutritional, or medical condition of individuals is expressly prohibited by law. 42 U.S.C. § 263a (2006); 42 C.F.R. § 493.3 (2006); COMAR 10.10.01.02 (2006). Genox is not a CLIA certified laboratory.

ELISA Testing Kits
Genox Corporation has
developed biomarker
assays to measure
several oxidative
damage biomarkers.

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).

The ‘Anti-4-Hydroxy-2-Nonenal 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.

Note: Genox’s Products are for RESEARCH USE ONLY. Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS.

Note: Genox’s Products are for RESEARCH USE ONLY.
Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS.