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Poly(deoxyinosinic-deoxycytidylic) acid sodium salt

聚(脱氧肌苷酸 - 脱氧胞苷酸)钠盐

Company: Sigma-Aldrich
Catalog#: P4929
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Electrophoresis Mobility Shift Assay
Author:
Date:
2014-04-05
[Abstract]  Protein (transcription factors and/or transcription cofactors)-binding to DNA is a critical event in regulation of transcription. Electrophoresis Mobility Shift Assay (EMSA), also known as gel shift assay, is a useful tool to detect protein- or protein complex-DNA/RNA interaction and to evaluate DNA binding specificity of transcription factors in vitro. Here we describe a simple method for EMSA with fluorescent dye-bound oligo DNA probes and recombinant protein expressed in bacterial cells. Using fluorescent dye instead of radioisotope enables easy handling and long-term storage of labelled-probes without reduction of detection sensitivity. [摘要]  与DNA结合的蛋白质(转录因子和/或转录辅因子)是调节转录的关键事件。 电泳迁移率变动分析(EMSA),也称为凝胶移位分析,是检测蛋白质或蛋白质复合物-DNA/RNA相互作用和评价转录因子的体外DNA结合特异性的有用工具。 在这里我们介绍一种简单的方法与荧光染料绑定寡DNA探针和表达在细菌细胞中的重组蛋白的EMSA。 使用荧光染料代替放射性同位素使得容易处理和长期储存标记的探针而不降低检测灵敏度。

EMSA Analysis of DNA Binding By Rgg Proteins
Author:
Date:
2013-08-05
[Abstract]  In bacteria, interaction of various proteins with DNA is essential for the regulation of specific target gene expression. Electrophoretic mobility shift assay (EMSA) is an in vitro approach allowing for the visualization of these protein-DNA interactions. Rgg proteins comprise a family of transcriptional regulators widespread among the Firmicutes. Some of these proteins function independently to regulate target gene expression, while others have now been demonstrated to function as effectors of cell-to-cell communication, having regulatory activitiesthat that are modulated via direct interaction with small signaling peptides. EMSA analysis can be used to assess DNA binding of either type of Rgg protein. EMSA analysis of Rgg protein activity has facilitated in vitro ... [摘要]  在细菌中,各种蛋白质与DNA的相互作用对于调节特异性靶基因表达是必需的。电泳迁移率变动分析(EMSA)是允许这些蛋白质-DNA相互作用的可视化的体外方法。 Rgg蛋白包括在坚牢菌中广泛的转录调节子家族。这些蛋白质中的一些独立地起作用以调节靶基因表达,而其它蛋白质已经被证明作为细胞与细胞间通信的效应物起作用,具有通过与小信号肽的直接相互作用调节的调节活性。 EMSA分析可用于评估任一类型的Rgg蛋白的DNA结合。 EMSG对Rgg蛋白活性的分析促进了体外证实调节靶标,通过DNA探针诱变鉴定精确的DNA结合位点,以及表征一些同源信号肽调节Rgg蛋白功能的机制(<例如在某些情况下中断DNA结合)。

A General EMSA (Gel-shift) Protocol
Author:
Date:
2011-02-05
[Abstract]  An electrophoretic mobility shift assay (EMSA), also referred to as mobility shift electrophoresis, a gel shift assay, gel mobility shift assay, band shift assay, or gel retardation assay, is a common technique used to study protein-DNA or protein-RNA interactions. The control lane (the DNA/RNA probe without protein present) will contain a single band corresponding to the unbound DNA or RNA fragment. If the protein is capable of binding to the fragment, the lane with protein present will contain another band that represents the larger, less mobile complex of nucleic acid probe bound to the protein, which is 'shifted' up on the gel (since it has moved more slowly). Here, a protocol to carry out an EMSA assay is described. [摘要]  电泳迁移率变动测定(EMSA),也称为迁移率变动电泳,凝胶迁移测定,凝胶迁移率变动测定,带移测定或凝胶阻滞测定是用于研究蛋白质-DNA或蛋白质-RNA的常见技术 互动。 对照泳道(不含蛋白质的DNA/RNA探针)将含有对应于未结合的DNA或RNA片段的单个条带。 如果蛋白质能够结合片段,存在蛋白质的泳道将包含另一条带,其代表与蛋白质结合的核酸探针的较大的,较少移动的复合物,其在凝胶上"移位"(因为它具有 移动更慢)。 这里,描述了进行EMSA测定的方案。

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