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使用抗茄碱单克隆抗体通过薄层色谱免疫染色法简单测定甾体生物碱苷。

A simple determination of steroidal alkaloid glycosides by thin-layer chromatography immunostaining using monoclonal antibody against solamargine.

作者信息

Tanaka H, Putalun W, Tsuzaki C, Shoyama Y

机构信息

Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka, Japan.

出版信息

FEBS Lett. 1997 Mar 10;404(2-3):279-82. doi: 10.1016/s0014-5793(97)00106-3.

Abstract

A method of determination for solasodine glycosides by using thin-layer chromatography (TLC) immunostaining was investigated. Solasodine glycosides separated by silica gel TLC were transferred to a polyvinylidene difluoride membrane. The membrane was treated with sodium periodate solution followed by bovine serum albumin (BSA), resulting in a solasodine glycoside-BSA conjugate. Conjugation was confirmed by matrix-assisted laser desorption/ionization mass spectrometry. Individual spots were stained by monoclonal antibody against solamargine. Immunostaining of solasodine glycosides was more sensitive compared to other stainings. The newly established immunostaining method can be extended to analysis of the distribution of solasodine glycoside in the plant body.

摘要

研究了一种采用薄层色谱(TLC)免疫染色法测定茄解定糖苷的方法。通过硅胶TLC分离得到的茄解定糖苷被转移到聚偏二氟乙烯膜上。该膜先用高碘酸钠溶液处理,然后用牛血清白蛋白(BSA)处理,得到茄解定糖苷 - BSA偶联物。通过基质辅助激光解吸/电离质谱法确认了偶联。用抗澳洲茄边碱单克隆抗体对各个斑点进行染色。与其他染色方法相比,茄解定糖苷的免疫染色更灵敏。新建立的免疫染色方法可扩展到分析茄解定糖苷在植物体内的分布。

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