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ANTI-FLAG M2 Affinity Gel

ANTI-FLAG ® M2亲和凝胶

Company: Sigma-Aldrich
Catalog#: A2220
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Biochemical Analysis of Caspase-8-dependent Proteolysis of IRF3 in Virus-infected Cells
Author:
Date:
2016-11-20
[Abstract]  Interferon regulatory factor 3 (IRF3) is a transcription factor, which is critical for the antiviral response against a wide range of viruses (Hiscott, 2007; Ikushima et al., 2013). It gets activated in virus-infected cells via Toll like receptors (TLRs), RIG-I (retinoic acid inducible gene 1) like receptors (RLRs), cyclic GMP-AMP synthase (cGAS) – stimulator of interferon genes (STING), which are sensors of viral components in the cells (Chattopadhyay and Sen, 2014a; 2014b; Hiscott, 2007). IRF3 is a cytoplasmic protein, upon activation by virally activated sensors it gets phosphorylated, translocated to the nucleus and binds to the interferon-sensitive response element (ISRE) of the gene promoters to induce their transcription (Hiscott, 2007). IRF3 has other functions, including ... [摘要]  干扰素调节因子3(IRF3)是一种转录因子,对于广泛病毒的抗病毒反应至关重要(Hiscott,2007; Ikushima等,2013)。通过Toll样受体(TLR),RIG-I(视黄酸诱导型基因1)受体(RLR),环状GMP-AMP合成酶(cGAS) - 干扰素基因刺激剂(STING),其在病毒感染的细胞中被激活,其中是细胞中病毒组分的传感器(Chattopadhyay和Sen,2014a; 2014b; Hiscott,2007)。 IRF3是一种细胞质蛋白,当被病毒激活的传感器激活时,其被磷酸化,转移到细胞核并与基因启动子的干扰素敏感反应元件(ISRE)结合以诱导其转录(Hiscott,2007)。 IRF3具有其他功能,包括直接刺激病毒感染细胞凋亡。在该途径中,不需要IRF3的转录活性(Chattopadhyay等,2013b; Chattopadhyay等,2016; Chattopadhyay等,2010; Chattopadhyay和Sen,2010; Chattopadhyay等,2011)。这些途径受宿主因素以及病毒的负调控。我们的研究表明IRF3可以通过caspase-8依赖性切割进行蛋白水解加工(Sears等,2011)。 ...

Chromatin Immunoprecipitation (ChIP) Assay for Detecting Direct and Indirect Protein – DNA Interactions in Magnaporthe oryzae
Author:
Date:
2015-11-05
[Abstract]  Chromatin immunoprecipitation (ChIP) is a powerful technology for analyzing protein-DNA interactions in cells. Robust ChIP procedures have been established for investigating direct interactions between protein and DNA. However, detecting indirect protein-DNA interactions in vivo is challenging. Recently, we used ChIP to analyze an indirect protein-DNA interaction between a putative histone demethylase, MoJmjC, and the promoter of the superoxide dismutase 1-encoding gene MoSOD1 in the rice blast fungus Magnaporthe oryzae (M. oryzae) (Fernandez et al., 2014). We tagged MoJmjC with the 3x FLAG epitope (Fernandez et al., 2014), instead of the larger and more commonly used GFP epitope, to mitigate against steric hindrance. We also employed ... [摘要]  染色质免疫沉淀(ChIP)是一种强大的技术,用于分析细胞中的蛋白质-DNA相互作用。已建立了稳健的ChIP程序用于研究蛋白质和DNA之间的直接相互作用。然而,在体内检测间接蛋白-DNA相互作用是具有挑战性的。最近,我们使用ChIP来分析推定的组蛋白去甲基化酶MoJmjC和在稻瘟病真菌Magnaporthe oryzae中超氧化物歧化酶1编码基因MoSOD1的启动子之间的间接蛋白质-DNA相互作用( oryzae )(Fernandez ,,2014)。我们用3x FLAG表位标记MoJmjC(Fernandez等人,2014),而不是更大和更常用的GFP表位,以减轻空间位阻。我们还采用了使用DSG和甲醛的两步交联策略,而不是更常用于分析直接蛋白质-DNA相互作用的一步甲醛交联方法,以便更好地捕获间接的MoJm - MoSOD1 DNA相互作用。此外,我们已经表明两步交联适用于GATA转录因子,Asd4及其同源结合位点之间的直接蛋白-DNA相互作用的ChIP分析(Marroquin-Guzman和Wilson,2015)。在这里,我们提供详细的协议的染色质免疫沉淀,与通用的两步交联,在M。 oryzae 。

Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
Author:
Date:
2015-07-05
[Abstract]  Nicotiana benthamiana (N. benthamiana) is a useful model system to transiently express protein at high level. This protocol describes in detail how to transiently express protein in N. benthamiana and how to carry out protein immunoprecipitation in this expression system. This protocol can be broadly used for investigation on protein-protein interaction, protein purification and other related protein assay. [摘要]  本塞姆氏烟草 ( )是一种用于高水平瞬时表达蛋白质的有用的模型系统。 该协议详细描述了如何在N中瞬时表达蛋白质。 本bent和如何在此表达系统中进行蛋白质免疫沉淀。 这个协议可以广泛用于调查蛋白质 - 蛋白质相互作用,蛋白质纯化和其他相关的蛋白质测定。

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