| Immunocytochemical Detection of Recombinant Biomphalysin on Schistosoma mansoni Sporocysts
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Author:
Date:
2013-11-20
[Abstract] Schistosomiasis, or bilharzia, is a tropical disease caused by worms of the genus Schistosoma which infect about 200 million people. The life cycle of the parasite requires Biomphalaria, a specific genus of freshwater snails, as intermediate. Using an interactome approach employing B. glabrata plasma and S. mansoni primary sporocyst extracts, we identified a new cytolytic protein called Biomphalysin that displays similarities to members of the β-PFT superfamily known to form channels in targeted membranes. To investigate its mechanism of action, we produced ...
[摘要] 血吸虫病或鼠伤寒沙菌是由感染约2亿人的蠕虫属引起的热带病。 寄生虫的生命周期需要作为中间体的淡水蜗牛的特定属的生物疟原虫。 使用采用em的interactome方法。 glabrata plasma和 我们鉴定了一种称为生物溶解素的新的溶细胞蛋白，其显示与已知在靶膜中形成通道的β-PFT超家族成员的相似性。 为了调查其作用机制，我们产生侧翼有N端6组氨酸标签的重组蛋白。 然后，我们调查了Biomphalysin与孢子囊体积相互作用的能力。 这个优化的协议描述免疫细胞化学程序检测组氨酸标记的重组蛋白在孢子被膜上。
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| Cell Proliferation Assay by Flow Cytometry (BrdU and PI Staining)
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Author:
Date:
2012-04-05
[Abstract] Cell Proliferation assays include an important set of fluorescence-based tests that can monitor cell health and cell division by evaluating DNA synthesis through thymidine incorporation. Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU is commonly used in the detection of proliferating cells in living tissues. BrdU can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication. Antibodies specific for BrdU can then be used to ...
[摘要] 细胞增殖测试是一个重要的基于荧光检测的实验，可以监测细胞健康和通过胸苷掺合来估量DNA合成从而监测细胞分裂。溴脱氧尿苷(5-溴代-2'-脱氧尿苷, BrdU) 是一种合成的核苷酸即胸苷的类似物. BrdU通常被用于检测活组织中再生的细胞。 BrdU可以掺合到复制细胞中新的合成的DNA中去（在细胞周期的S期），在DNA复制是代替胸苷。. BrdU 特异性抗体可以用于检测这个参入的化学品，从而检测出细胞是否在复制其DNA。对抗体的结合需要DNA变形，通常将细胞置于酸性环境或者加热。掺合的BrdU通常通过标记抗BrdU的抗体和DNA染料碘化丙啶（PI）而用流式细胞仪检测来实现对细胞周期的检测。
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| Cell Cycle Analysis using Propidium Iodide Staining with GFP Detection
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Author:
Date:
2012-04-05
[Abstract] Infecting mammalian cells with a GFP construct to overexpress or knockdown target genes is one of the most commonly used methods to study and manipulate gene expression. To determine the target gene function on the cell cycle, analyzing the cell cycle (propidium iodide, PI staining) of GFP positive cells vs GFP negative cells is needed. Usually simple fixation of cells with 70% EtOH for PI staining tends to quench GFP signal; paraformaldehyde (PFA) fixation before ETOH fixation could help to sustain the GFP signal.
[摘要] 用一个GFP标记的质粒转染哺乳动物细胞过表达或者调低目的基因是一种最常用的操纵基因表达的方法。想要在细胞循环中检测目的基因的功能，分析GFP阳性细胞和GFP阴性细胞的细胞循环（碘化丙啶，PI染色）是必要的。通常为PI染色用70 %乙醇简单固定细胞淬火GFP信号，在乙醇固定前用低聚甲醛（PFA）固定能帮助染GFP信号。
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