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DDSA (Dodecenyl Succinic Anhydride) EM grade redistilled 100g

DDSA(十二烯基琥珀酸酐)EM级再蒸馏100g

Company: Agar Scientific
Catalog#: AGR1053
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Immuno-EM Analysis of PF13_0191-GFP Expressing Parasites
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2014-06-05
[Abstract]  This protocol was used to prepare pre-embedding samples of Plasmodium falciparum blood stage parasites that overexpressed the parasite protein PF13_0191 tagged with GFP. Using GFP-specific antibodies and Protein A-Gold the localisation of the overexpressed protein in the infected host cell was determined using standard transmission electron microscopy (EM). Pre-embedding EM is a common method where the antibodies are introduced before embedding and sectioning. This method avoids the problem that antigens are often difficult to detect on EM-sections after embedding. In the method described here antigens in the parasite-infected host cell are detected. Entry of the antibody is made possible through permeabilisation of the host cell with tetanolysin. In principle this method could ... [摘要]  该方案用于制备过表达用GFP标记的寄生虫蛋白PF13_0191的恶性疟原虫血液阶段寄生虫的预嵌入样品。使用GFP特异性抗体和蛋白A-Gold,使用标准透射电子显微镜(EM)测定过表达的蛋白在感染的宿主细胞中的定位。预嵌入EM是其中在包埋和切片之前引入抗体的常见方法。该方法避免了嵌入后在EM-切片上通常难以检测抗原的问题。在本文所述的方法中,检测寄生虫感染的宿主细胞中的抗原。通过宿主细胞用破伤风溶蛋白透化可以进入抗体。原则上,如果样品在添加相关抗体之前被适当地固定和透化,则该方法也可以用于检测寄生虫内的抗原。虽然抗体的获取将避免嵌入后方法中经常见到的检测问题,但是该方法将产生相对较差的形态。

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