Analysis of host-cell responses by immunoblotting, ELISA, and real-time PCR.
Analysis of host-cell responses by immunoblotting, ELISA, and real-time PCR.
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通过免疫印迹、ELISA 和实时 PCR 分析宿主细胞反应。
DOI：
10.1007/978-1-61779-539-8_23
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Moyes DL
中科院分区：
文献类型：
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作者：
Moyes DL
细胞内对外部病原体/刺激的反应对于宿主对感染的反应至关重要。用于分析这些答复的方法分为许多类别。可以使用常规免疫印迹技术或免疫沉淀来筛选作为信号级联的一部分的蛋白质的活化，以鉴定修饰的蛋白质或蛋白质复合物的存在。可以通过EMSA或ELISA筛选转录因子活性以鉴定这些因子的DNA结合。最后，可以使用实时PCR方法定量活化基因的表达。在这里，我们将展示如何执行这些检测，并讨论每一个的相对优点。
Intracellular responses to external pathogens/stimuli are crucial to the host’s response to infection. The methods used to analyse these responses fall into many categories. Activation of proteins as part of a signal cascade can be screened for using conventional immunoblotting techniques or immunoprecipitation to identify the presence of modified proteins or protein complexes. Transcription factor activity can be screened for by EMSA or ELISAs to identify DNA binding of these factors. Finally, expression of activated genes can be quantified using real-time PCR methods. Here, we will show how to perform these assays and discuss the relative merits of each.