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1-StepTM NBT/BCIP Substrate Solution

1-Step TM NBT / BCIP底物溶液

Company: Thermo Fisher Scientific
Catalog#: 34042
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Colony Immunoblotting Assay for Detection of Bacterial Cell-surface or Extracellular Proteins
Author:
Date:
2013-09-05
[Abstract]  This simple protocol describes how to detect antigens from agar-grown bacterial colonies transferred to nitrocellulose using specific antibodies. The protocol is well suitable for detection of bacterial proteins exposed on the cell surface or secreted to the extracellular space and it can be modified also for detection of intracellular proteins. The assay can distinguish bacterial clones with different expression rates (high, medium and low) from colonies that do not express target protein. We used this assay for screening of Mat fimbriae-producing Escherichia coli mutants obtained by mini-Tn5 transposon mutagenesis and immunomagnetic separation (Lehti et al., 2013). [摘要]  这个简单的协议描述如何使用特异性抗体检测从转移到硝酸纤维素的琼脂生长的细菌菌落的抗原。 该方案非常适合于检测暴露于细胞表面或分泌至细胞外空间的细菌蛋白,并且其还可以被修饰以用于检测细胞内蛋白。 该测定可以区分具有不同表达率(高,中和低)的细菌克隆与不表达靶蛋白的集落。 我们使用该测定法用于筛选通过mini-Tn5转座子诱变和免疫磁性分离获得的产生Mat菌毛的大肠杆菌突变体(Lehti等人,2013)。

Western Analysis of Histone Modifications (Aspergillus nidulans)
Author:
Date:
2013-04-05
[Abstract]  Western blotting allows for the specific detection of proteins and/or modifications of proteins by an antibody of interest. This protocol utilizes a crude nuclei extraction protocol for Aspergillus nidulans to enrich for histones and other nuclear proteins prior to gel electrophoresis. Post translational modifications of histones may then be easily detected. After electrophoresis, the selected antibodies are used to detect and quantify levels of the modifications of interest. [摘要]  蛋白质印迹允许由感兴趣的抗体特异性检测蛋白质和/或蛋白质的修饰。 该方案在凝胶电泳之前利用构巢曲霉的粗核提取方案来富集组蛋白和其它核蛋白。 然后可以容易地检测组蛋白的翻译后修饰。 电泳后,所选择的抗体用于检测和定量感兴趣的修饰的水平。

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