Higashi H, Fukui Y, Ueda S, Kato S, Hirabayashi Y, Matsumoto M, Naiki M
J Biochem. 1984 May;95(5):1517-20. doi: 10.1093/oxfordjournals.jbchem.a134760.
A sensitive enzyme-immunochemical staining method was developed for detection of N-glycolylneuraminic acid (NeuGc)-containing glycosphingolipids (GSLs) on silica gel thin-layer chromatography. The procedure consists of immune reaction among NeuGc-containing GSLs, affinity-purified chicken anti-NeuGc-LacCer and horseradish peroxidase-conjugated rabbit anti-chicken IgG, and the peroxidase reaction using 4-chloro-1-naphthol as a chromogenic substrate. Quantitative determination was achieved by direct densitometric scanning of the enzyme-immunostained spots on the chromatogram. As little as 0.5 pmol of NeuGc-LacCer, NeuGc-nLcOse4Cer, and NeuGc-nLcOse6Cer (0.64-1.0 ng) could be detected with a good signal-to-noise ratio. A semi-linear detector response was observed up to 50 pmol of each GSL. This procedure can be applied easily to other glycolipid antigen systems.
开发了一种灵敏的酶免疫化学染色方法，用于在硅胶薄层色谱上检测含N-羟乙酰神经氨酸（NeuGc）的糖鞘脂（GSL）。该程序包括含NeuGc的GSL、亲和纯化的鸡抗NeuGc-LacCer和辣根过氧化物酶偶联的兔抗鸡IgG之间的免疫反应，以及使用4-氯-1-萘酚作为显色底物的过氧化物酶反应。通过对色谱图上酶免疫染色斑点进行直接光密度扫描进行定量测定。低至0.5 pmol的NeuGc-LacCer、NeuGc-nLcOse4Cer和NeuGc-nLcOse6Cer（0.64-1.0 ng）就能以良好的信噪比被检测到。每种GSL在高达50 pmol时观察到半线性检测器响应。该程序可轻松应用于其他糖脂抗原系统。
