4-羟基壬醛
脂质过氧化
化学
单克隆抗体
生物化学
氧化应激
组氨酸
结合
分子生物学
酶
抗体
生物
免疫学
数学分析
数学
作者
Suzana Borović,Filip Rabuzin,Georg Waeg,Neven Žarković
标识
DOI:10.1080/10715760600693422
摘要
Highly reactive aldehyde 4-hydroxynonenal (HNE) is the final product of lipid peroxidation, known as a second messenger of free radicals and a signaling molecule. It forms protein conjugates involved in pathology of various diseases. To determine cellular HNE-protein conjugates we developed indirect ELISA based on well-known, monoclonal antibody against HNE-histidine (HNE-His) adducts. The method was calibrated using HNE-albumin conjugates as standards (R(2) = 0.999) and validated on human osteosarcoma cell cultures (HOS). The ELISA showed good sensitivity (8.1 pmol HNE-His/mg of protein), precision ( +/- 8% intra-assay and +/- 12% inter-assay) and spiking recovery ( +/- 9%). The assay revealed 60-fold increase of cellular HNE-His adducts upon copper-induced lipid peroxidation of HOS. The ELISA matched HNE-immunocytochemistry of HNE-treated HOS cells and quantified the increase of cellular HNE-His conjugates in parallel to the decrease of free HNE in culture medium. The ELISA was developed as ELISA Stress for severe lipid peroxidation and ELISA Fine for studies on HNE physiology.
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