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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Novel chemiluminescent Western blot blocking and antibody incubation solution for enhanced antibody‐antigen interaction and increased specificity
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Novel chemiluminescent Western blot blocking and antibody incubation solution for enhanced antibody‐antigen interaction and increased specificity

机译：新型化学发光蛋白质印迹阻断和抗体培养溶液，用于增强抗体 - 抗原相互作用和增加的特异性

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摘要

Abstract Western blotting is a ubiquitous tool used in protein and molecular biology research, providing information about the presence, size, relative abundance, and state of a protein in a mixture. First, the proteins in a sample are separated by size using SDS‐PAGE then transferred onto a membrane for detection with a set of primary and secondary antibodies. High‐quality Western data requires high signal‐to‐noise ratios, which depend upon reduction of nonspecific antibody interactions. Blocking is a critical step in the Western blot method as it prevents the antibodies from binding nonspecifically to the membrane and irrelevant proteins. A solution of nonfat dry milk (NFDM) in physiological buffer is commonly used for this purpose, but does not perform well with every type of antibody and is not optimal for low‐abundance proteins. We present a novel blocking solution for chemiluminescent Western blots, AdvanBlock?‐chemi, which outperforms NFDM in experiments with 20 unique antibodies by increasing signal‐to‐noise ratios and minimizing nonspecific binding. This solution enhances protein detection by Western blot and provides consistent results for detection of low abundant and modified proteins.
机译：摘要Western Blotting是一种用于蛋白质和分子生物学研究的普遍存在的工具，提供有关混合物中蛋白质的存在，大小，相对丰度和状态的信息。首先，使用SDS-PAGE将样品中的蛋白质通过尺寸分离，然后将其转移到膜上以用一组初级和二抗检测。高质量的西方数据需要高信噪比，这取决于减少非特异性抗体相互作用。阻断是蛋白质印迹方法中的关键步骤，因为它可以防止抗体非特异性地结合到膜和不相关的蛋白质中。生理缓冲液中的非含量干乳（NFDM）的溶液通常用于此目的，但对每种类型的抗体并不顺利，对低丰度蛋白不是最佳的。我们提出了一种用于化学发光蛋白质印迹的新型封闭溶液，Advanblockα-Chemi，其在通过增加信噪比并最小化非特异性结合，以20个独特的抗体的实验表现出NFDM。该溶液通过Western印迹增强蛋白质检测，并提供了检测低丰富和改性蛋白质的一致结果。

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