| 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 pyrenoid; Yamano et al., 2010), LCI1 (plasma membrane-localized inorganic carbon transporter; Ohnishi et al., 2010), HLA3 (plasma membrane-localized ABC-type bicarbonate transporter; Yamano et ...
[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 cell level. With detection levels that can be as low as one cell in 100,000, the ELISpot is one of the most sensitive cellular assays available. Depending on the substance analyzed, the ELISpot assay ...
[摘要] 119.txt
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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的重要信号分子，并且在许多人肿瘤中被组成型激活，并且具有致癌潜力和抗凋亡活性。 STAT3被Tyr705处的磷酸化激活，其诱导二聚化，核转位和DNA结合。 蛋白质印迹最常用于通过使用对磷酸化酪氨酸特异的抗体来检测STAT3的活化。
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