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Lead(II) citrate tribasic trihydrate


Company: Sigma-Aldrich
Catalog#: 15326
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Immunogold Electron Microscopy of the Autophagosome Marker LC3
[Abstract]  Even though autophagy was firstly observed by transmission electron microscopy already in the 1950s (reviewed in Eskelinen et al., 2011), nowadays this technique remains one of the most powerful systems to monitor autophagic processes. The autophagosome, an LC3-positive double membrane structures enclosing cellular materials, represents the key organelle in autophagy and its simple visualization and/or numeration allow to draw important conclusions about the autophagic flux. Therefore, the accurate identification of autophagosomes is crucial for a comprehensive and detailed dissection of autophagy. Here we present a simple protocol to identify autophagosomes by transmission electron microscopy coupled to immunogold labeling of LC3 starting from a relatively low cell number, which ... [摘要]  尽管早在20世纪50年代就已经通过透射电子显微镜观察了自噬(在Eskelinen等人2011年的综述中),但是现在这种技术仍然是监测自噬过程的最强大的系统之一。 自噬体是包含细胞物质的LC3阳性双层膜结构,代表了自噬的关键细胞器,其简单的可视化和/或计数允许得出关于自噬流的重要结论。 因此,准确鉴定自噬体对自噬的全面和详细的分析至关重要。 在这里我们提出一个简单的协议,以确定autophagosomes透射电子显微镜耦合LC3的免疫金标记从一个相对较低的细胞数量,我们最近开发遵循病毒介导的人类癌变期间的自噬途径。

【背景】自噬体代表了macroautophagy的关键结构,这是一种细胞胞质成分的分解代谢系统。巨自噬(或简单地自吞噬)由吞噬细胞的形成引发,所述吞噬细胞能够自身扩张吞噬细胞器和蛋白质,所述蛋白质最终闭合在螯合成分周围形成被称为自噬体的细胞器。接下来,在成熟过程中,自噬体可以与溶酶体融合以形成自溶酶体,其中被捕获的物质被位于溶酶体限制性膜中的泵降解并再循环回(Glick et al。,2010)。

由于自噬功能障碍与各种人类疾病,病毒感染,神经退行性疾病,免疫功能和癌症有关(Schneider and ...

Immunoelectron Microscopy in Chlamydomonas Cells
[Abstract]  The method of immunoelectron microscopy is intended for localization of proteins inside the cells of Chlamydomonas reinhardtii or other microalgae and cyanobacteria. This protocol was used to study localization of carbonic anhydrase Cah3 with antibodies raised in rabbit, though it can be used to localize any other abundant protein. Primary rabbit antibodies are recommended because they react quickly and specifically with proteins of C. reinhardtii. If primary antibodies other than rabbit are used, the blocking procedure and time of incubation with primary and secondary antibodies should be adjusted. [摘要]  免疫电子显微镜法的目的是将蛋白质定位在莱茵衣藻或其它微藻和蓝细菌的细胞内。 该方案用于研究碳酸酐酶Cah3与在兔中产生的抗体的定位,尽管其可以用于定位任何其他丰富的蛋白质。 建议使用初级兔抗体,因为它们能快速和特异性地与C的蛋白反应。 reinhardtii 。 如果使用除兔以外的第一抗体,应当调整与第一和第二抗体孵育的封闭程序和时间。