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HEL may be a useful biomarker for initial stage of lipid peroxidation

Hexanoyl-Lys (HEL) ELISA Kit and related product

Background

Oxidative damage of lipids caused by reactive oxygen species (ROS) play an important role in some diseases, lesion of cell functions and aging. Aldehydessuch 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. HEL may be a useful biomarker for initial stage of lipid peroxidation.Monoclonal antibodies and ELISA kit have been developped, and HEL can be detected in oxidatively modified LDL, in human atherosclerotic lesions, human urine and serum. It is also reported that HELis formed in rat muscle during exercise, and the formation is prohibited by antioxidants such as flavonoids.
HEL is formed by the reaction of linoleic acid hydroperoxide and Lysine, and a biomarker for oxidative stress. HEL ELISA kit is a competitive enzyme-linked immunosorbent assay for quantitative measurement of hexanoyl-Lys adduct. This kit is based on the monoclonal antibody clone 5H4, which is specific for HEL. Suitable for urine, serum and other biological samples.

Principle

Assay Procedure

Kit component

1. HEL Microtiter Plate Precoated with HEL (8x12 wells, Split Type) 1 Plate
2. Primary Antibody Monoclonal Antibody specific for HEL 1 Vial (7ml)
3. Secondary Antibody HRP-Conjugated Anti Mouse IgG Antibody 1 Vial
4. Secondary Antibody Buffer Phosphate Buffered Saline 1 Vial (12ml)
5. Chromogen 3,3',5,5'-Tetramethylbenzidine 1 Vial (250 µL)
6. Chromogen Buffer Hydrogen Peroxide/ Citrate-Phosphate Buffer 1 Vial (12ml)
7. Washing Buffer (5X) Concentrated Phosphate Buffered Saline 2 Vials (25mL X 2)
8. Stop Solution 1M Phosphoric acid 1 Vial (12mL)
9. *Standard A-F Bz-Gly-Hexanoyl-Lys 1 Vial each(500µL)
10. Plate Seal Adhesive seal to prevent evaporation 2 sheets

*(A: 2.6, B:7.7, C:22.7, D:69.7, E:207, F:624 nmol/L)

Standard curve example

Product List

Product Name Cat# Quantity Price

Hexanoyl-Lys(HEL) ELISA kit

NNS-KHL-700E-EX 96WELL

¥ 76,800
$ 1024
€ 768

References
  •  Kato Y. et al., Formation of Nepsilon-(hexanonyl)lysine in protein exposed to lipid hydroperoxide. A plausible marker for lipid hydroperoxide-derived protein modification. J Biol Chem. 1999 Jul 16;274(29):20406-14. PMID: 10400665
  •  Kato Y. et al., 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. 2000 Aug 2;274(2):389-93. PMID: 10913348
  •  Ryo K. et al, Possible involvement of oxidative stress in salivary gland of patients with Sjogren's syndrome. Pathobiology. 2006;73(5):252-60. PMID: 17314496
  •  Suzuki T. et al, Effect of prophylactically administered edaravone during antegrade cerebral perfusion in a canine model of old cerebral infarction. J Thorac Cardiovasc Surg. 2007 Mar;133(3):710-6. PMID: 17320569
  •  Shimizu K. et al, Increased serum levels of N(epsilon)-(hexanoyl)lysine, a new marker of oxidative stress, in systemic sclerosis. J Rheumatol. 2008 Nov;35(11):2214-9. Epub 2008 Sep 1. PMID: 18785309
  •  Tamura H. et al, Oxidative stress impairs oocyte quality and melatonin protects oocytes from free radical damage and improves fertilization rate. J Pineal Res. 2008 Apr;44(3):280-7. doi: 10.1111/j.1600-079X.2007.00524.x. PMID: 18339123
  •  Sakamoto Y. et al, The assessment of oxidative stress in infertile patients with varicocele. BJU Int. 2008 Jun;101(12):1547-52. doi: 10.1111/j.1464-410X.2008.07517.x. Epub 2008 Feb 21. PMID: 18294306
  •  Kageyama Y. et al, Etanercept reduces the oxidative stress marker levels in patients with rheumatoid arthritis. Rheumatol Int. 2008 Jan;28(3):245-51. Epub 2007 Jul 28. PMID: 17661050
  •  Rummenie VT. et al, Tear cytokine and ocular surface alterations following brief passive cigarette smoke exposure. Cytokine. 2008 Aug;43(2):200-8. doi: 10.1016/j.cyto.2008.05.011. Epub 2008 Jul 3. PMID: 18602273
  •  Sakai K. et al, Determination of HEL (Hexanoyl-lysine adduct): a novel biomarker for omega-6 PUFA oxidation. Subcell Biochem. 2014;77:61-72. doi: 10.1007/978-94-007-7920-4_5. PMID: 24374918