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Basic Methods in Cellular and Molecular Biology
Protein Detection by Western Blot
Description
Western blotting is a lab method used to detect specific proteins in samples that have already been separated by electrophoresis. It helps scientists find a single protein within a complex mixture by using antibodies that recognize it.
First, the proteins are separated by SDS-PAGE, which stands f...
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Transcript
蛋白免疫印迹杂交是被许多科研人员用来通过抗体鉴定电泳分离样品中是否存在某特定蛋白的强大技术。
该技术的完成需要三个基本步骤:电泳印迹转移、免疫印迹杂交和结果检测。在这些步骤之前，先要进行聚丙酰胺凝胶电泳，它根据蛋白分子大小将变性蛋白进行分离。
电泳印迹转移，也称凝胶转移，要用到转移夹盒来固定"三明治"，和一个将蛋白质从聚丙酰胺凝胶转移到膜上的装置。电泳转移三明治包含:凝胶和特殊的膜，它们被夹在两张滤纸中间。转移过程中施加的电场将使得蛋白质穿过凝胶，在电荷和疏水作用的帮助下结合到薄膜上。
免疫印迹杂交是使用抗体检测薄膜上的特定蛋白。抗体是大的Y型蛋白，有...
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