Marchini Barbara, De Nuccio Luca, Mazzei Mauro, Mariottini Gian Luigi
Dipartimento di Medicina Interna, Università di Pisa, Italy.
Riv Biol. 2004 Aug-Dec;97(3):505-15.
Nematocyst isolation from surrounding tissue is an important step to characterize Cnidarian venom. Although several protocols have been used to extract venoms from cnidarian tissues, the complete isolation of nematocysts from tissue is still difficult. The goal of the present work was to evaluate the effectiveness of three different media, Percoll, Ficoll and Methylcellulose in isolating nematocysts from Pelagia noctiluca tentacles by centrifugation. The complete sedimentation of nematocysts and tissue fragments to the bottom of the test tubes was observed in Ficoll and Methylcellulose suspensions. The best result was obtained using a discontinuous density gradient of Percoll: three types of nematocysts were concentrated in three different fractions along the density gradient. Protein assay and preliminary chromatographic analyses confirmed these results.
从周围组织中分离刺丝囊是鉴定刺胞动物毒液的重要步骤。尽管已经使用了几种方法从刺胞动物组织中提取毒液,但从组织中完全分离刺丝囊仍然很困难。本研究的目的是评估三种不同介质( Percoll、Ficoll和甲基纤维素)通过离心从夜光游水母触手中分离刺丝囊的有效性。在Ficoll和甲基纤维素悬浮液中观察到刺丝囊和组织碎片完全沉淀到试管底部。使用不连续密度梯度的Percoll获得了最佳结果:三种类型的刺丝囊沿密度梯度集中在三个不同的组分中。蛋白质测定和初步色谱分析证实了这些结果。