Genox

Anti-Hexanoyl-Lysine Monoclonal Antibody
(Anti HEL monoclonal antibody: clone 5H4) 
(Catalog #MHL-020P for 20 microgram vial)

Product Insert (pdf)pdf


What is Hexanaoyl-Lysine?

Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in lesion of cell functions and aging. Aldehydes such as malondi-aldehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) have been reported as one of the advanced lipid peroxidation products. In the earlier stages of lipid peroxidation,
13-hydroperoxyoctadecanoic acid (13-HPODE) is found to be covalently bound to proteins1). Hexanoyl-Lysine adduct  is a novel lipid hydroperoxide-modified lysine residue, formed by oxidative modification by oxidized omega-6 fatty acids such as linoleic acid or arachidonic acid. This monoclonal antibody (clone 5H4) is specific for proteins and peptides containing Hexanoyl-Lysine. Suitable for detection by immuno-histochemistry and western blotting.

mono2a

 

Source

:

Mouse

Class

:

IgG

Antigen

:

N epsilon hexanoyl-Keyhole limpet hemocyanin (HEL-KLH)

Specifity

:

Specific to Hexanoyl-Lys adducts.

Form

:

Lyophilized power. Containing 10mM PBS(pH7.4), 5% sucrose,
1% BSA and 0.05% Procline 950. 100 micro g/mL of IgG.

Application


:

Immunohistochemistry.
Recommended antibody concentration
is 2 micro gram/mL on paraformaldehyde fixed tissue.

Storage

:

Less than -20°C

Specificity of anti HEL antibody.

 


Detection of HEL at Human atherosclerotic lesions.
Photo: kindly provided by Dr. Naito

 

IMMUNOHISTOCHEMISTRY
1)

Paraffin sections were deparaffinized and rehydrated.

2)

Antigen retrieval by microwave for 5 minutes in 10mM citrate buffer (pH6.0).

3)

Sections were pretreated for 30 min with 3% non-immune animal serum in 1% BSA / PBS (pH7.4)..

4)

Sections were incubated overnight at 4°C with a antibody at a dilution of 2 micro gram/mL.

5)

Antibody binding was visualized by the avidin-biotin-immuno peroxidase complex method using the Vectastain ABC kit (Vector, Burlingame, CA) according to the manufacturer's instructions.

6)
3,3-Diaminobenzidine tetrahydrochloride was used as the chromogen.
7)

Hematoxylin or Eosin was used as the counter stain.


REFERENCES

1)
Formation of NfA-(hexanonyl) lysine in protein exposed to lipid hydroperoxide.
Y. Kato, Y. Mori,Y. Makino, Y. Morimitsu, S. Hiroi, T. Ishikawa, T. Osawa.
J. Biol. Chem. 274(29) p20406-20414 (1999).
2)
Detection of lipid hydroperoxide-derived protein modification with polyclonal antibodies.
Y. Kato, T. Osawa.
Methods in Molecular Biology, vol. 186, Oxidative stress biomarkers and antioxidant protocols, D. Armstrong, Ed., Human Press Inc., NJ, USA, p37-44.
3)
Preparation of monoclonal antibody to NfA-(hexanonyl) lysine: application to the evaluation of protective effects of flavonoid supplementation against exercise-induced oxidative stress in rat skeletal muscle.
Y. Kato, Y. Miyake, K. Yamamoto, Y. Shimomura, H. Ochi, Y. Mori, T. Osawa.
Biochem. Biophys. Res. Commun. 274(2), p389-393 (2000).
4)
The protective effects of tetrahydrocurcumin on oxidative stress in cholesterol-fed rabbits.
M. Naito, Wu X, H. Nomura, M. Kodama, Y. Kato, T. Osawa.
J. Atheroscler Thromb. 9(5), p243-250 (2002).

5)

Formation of Nepsilon-(hexanonyl) lysine in oxidized human very-low density lipoprotein.
H. Arai, Y. Kato, K. Fukunaga, S. Mohri, K. Nakamura.
J. Electrophoresis 48, pg37-40 (2004).

 

The ‘Anti-Hexanaoyl-Lysine 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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Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS. 

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