| Indirect Immunofluorescence Assay in Chlamydomonas reinhardtii
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Author:
Date:
2016-07-05
[Abstract] Determining the protein localization is essential to elucidate its in vivo function. Fluorescence-tagged proteins are widely used for it, but it is sometimes difficult to express tagged proteins in Chlamydomonas. Alternatively, indirect immunofluorescence assay is also one of the widely used methods and many reports determining the localization of Chlamydomonas proteins using this method are published. Here, we introduce a protocol of indirect immunofluorescence assay adapted from our papers reporting LCIB (CO2-recycling factor in the vicinity of ...
[摘要] 确定蛋白质定位是阐明其体内功能所必需的。荧光标记的蛋白质被广泛使用，但有时难以在衣原体中表达标记的蛋白质。或者，间接免疫荧光测定也是广泛使用的方法之一，并且公开了使用该方法确定衣藻蛋白定位的许多报道。在这里，我们介绍间接免疫荧光测定方案改编自我们的论文报告LCIB（在芘类化合物附近的CO 2 - 再循环因子; Yamano等人，2010）， LCI1（质膜定位的无机碳转运蛋白; Ohnishi等人，2010），HLA3（浆膜定位的ABC-型碳酸氢盐转运蛋白; Yamano等人，2015） ）和LCIA（叶绿体包膜阴离子通道; Yamano等人，2015）在莱茵衣藻（Chlamydomonas reinhardtii）中。这里描述的协议可以用于观察其他藻类细胞感兴趣的蛋白质。
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| INF-gamma Release ELISpot Assay
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Author:
Date:
2012-03-20
[Abstract] IFN-γ (Interferon-gamma) is produced mainly by activated T cells and NK cells. Production of IFN-γ (Interferon-gamma) by helper T cells as well as cytotoxic T cells is a hallmark of the TH1-type phenotype, thus, high-level production of IFN-γ (Interferon-gamma) is typically associated with effective host defense against intracellular pathogens. The Enzyme-Linked ImmunoSpot (ELISpot) assay is commonly used to assess the function of antigen specific T cells by detecting IFN-γ release. The ELISpot assay is a very sensitive immunoassay, allowing the detection of a secreted cytokine at the single ...
[摘要] IFN-γ（干扰素-γ）主要产生于激活的T细胞和NK细胞。TH1型响应的特点是通过辅助T细胞和细胞毒T细胞产生IFN-γ，所以高水平IFN-γ的产生是由于寄主应对细胞内的病原体产生了有效的防御。酶联免疫斑点法通常用于评估抗原特异的T细胞的功效，即检测IFN-γ的释放。酶联免疫斑点法是一种非常灵敏的免疫检测方法，它可以在单细胞水平检测细胞因子的分泌。酶联免疫斑点法是最灵敏的细胞检测方法之一，它检测的灵敏度可以达到1:100000个细胞。依赖于化学物质的分析，酶联免疫斑点分析可以比传统的ELISA灵敏20到200倍。
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| Western Blot for Detecting Phosphorylated STAT3
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Author:
Date:
2011-08-20
[Abstract] The STAT3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors and is constitutively activated in a number of human tumors and possesses oncogenic potential and anti-apoptotic activities. STAT3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding. Western blot is most commonly used to detect the activation of STAT3 by using an antibody that is specific for the phosphorylated tyrosine705.
[摘要] Stat3转录因子是许多细胞因子和生长因子受体中重要的信号分子，它在许多人类的肿瘤细胞中持续激活，具备潜在的致癌能力和抗凋亡能力。Stat3通过705位酪氨酸磷酸化被激活，进而诱导它的二聚化，核定位以及DNA结合功能。STAT3的活性经常通过能识别705位酪氨酸磷酸化的抗体来检测。
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