Japan Institute for the Control of Aging: JaICA Oxidative Stress
Anti Hexanoyl-lysine adduct antibody
(Anti HEL monoclonal antibody)
A new biomarker for lipid peroxidation
Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in some diseases, 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. But recently in the earlier stage of lipid peroxidation, 13-hydroperoxyoctadecanoic acid (13-HPODE) is found to be covalently bound to proteins1). Hexanoyl-Lysine adduct (HEL) is a novel lipid hydroperoxide-modified lysine residues. HEL is formed by oxidative modification by oxidized omega-6 fatty acids such as linoleic acid or arachidonic acid. This monoclonal antibody (clone 5H4)is spesific for proteins and peptides containing HEL. Suitable for HEL detection by immunohistochemistry and western blotting.

Cisplatin-treated rat kidney.
(Dr. Sugiyama, Tottori Univ.)

Detection of HEL at Human atherosclerotic lesions.
(Dr. Naito)
Product name Code Specifications Application NOTICE
Anti HEL antibody(clone 5F12) MHL-021P 20 µg of IgG IHC Current product
Anti HEL antibody(clone 5H4) MHL-020P 20 µg of IgG IHC Discontinued

Made in Japan.

The clone of anti HEL monoclonal antibody has been changed from 5H4 (code. MHL-020P) to 5F12 (code. MHL-021P). If you need old product, please contact our technical support.

Related products:    Hexanoyl-Lys ELISA kit   
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
Clone #: 5F12
Subclass: IgG1 kappa
Antigen: N epsilon hexanoyl-Keyhole limpet hemocyanin (HEL-KLH)
Specificity: Specific to Hexanoyl-Lys adducts.
Form: Lyophilized power. Containing 10mM PBS(pH7.4), 5% sucrose, 1% BSA and 0.05% Procline 950. 100 µg/mL of IgG.
Application: Immunohistochemistry
(Recommended antibody concentration is 2 µg/mL on paraformaldehyde fixed tissue), and ELISA
Storage: Less than -20°C
Fixation: Formalin fixation. Slides should be prepared 3 um thick.
Antigen retrieval is not needed, but in some cases antigen retrieval may improve staining.
Blocking: Apply normal rabbit serum (DAKO corporation, Kyoto, Japan) in 1% BSA/PBS for 30 minites to the specimens for the inhibition of non-specific binding of secondary antibody.
For blocking reagent, please use normal serum from the same species as second antibody used. For example, if you use biotin-labeled goat anti mouse IgG, normal goat serum may be suitable. If sample tissues are from mouse, blocking of internal mouse IgG will be needed. See technical information.
Anti HEL antibody: Apply anti HEL antibody (2 µg/mL) to the specimens, incubate overnight at 4 °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).
1) Yoji Kato, Yoko Mori, Yuko Makino, Yasujiro Morimitsu, Sadayuki Hiroi, Toshitsugu Ishikawa, Toshihiko Osawa, Formation of N epsilon-(hexanonyl) lysine in protein exposed to lipid hydroperoxide. J Biol Chem 274(29), p20406-20414 (1999)
Identification of HEL, which is lysine adduct of 13-HPODE.
2) Kato Y, Miyake Y, Yamamoto K, Shimomura Y, Ochi H, Mori Y, Osawa T, Preparation of a monoclonal antibody to N(epsilon)-(Hexanonyl)lysine: application to the evaluation of protective effects of flavonoid supplementation against exercise-induced oxidative stress in rat skeletal muscle. Biochem Biophys Res Commun 274(2),p389-393(2000)
Development and characterization of anti HEL monoclonal antibody.
3) Fukuchi Y, Miura Y, Nabeno Y, Kato Y, Osawa T, Naito M, Immunohistochemical detection of oxidative stress biomarkers, dityrosine and N(epsilon)-(hexanoyl)lysine, and C-reactive protein in rabbit atherosclerotic lesions. J Atheroscler Thromb 15(4)p185-192(2008)
Application to rabbit atherosclerotic lesions.
4) Maeda R, Noiri E, Isobe H, Homma T, Tanaka T, Negishi K, Doi K, Fujita T, Nakamura E, A water-soluble fullerene vesicle alleviates angiotensin II-induced oxidative stress in human umbilical venous endothelial cells. Hypertens Res 31(1)p141-151(2008)
Application to cultured cells (HUVECs).
5) Sango K, Yanagisawa H, Kato K, Kato N, Hirooka H, Watabe K: Differential Effects of High Glucose and Methylglyoxal on Viability and Polyol Metabolism in Immortalized Adult Mouse Schwann Cells. Open Diabetes J. 1, p1-11(2008)
Application to cultured cell from mouse.
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.