Antibody Purification from Western Blotting

Abstract

This protocol describes a method of purifying antibodies from sera with denatured antigens immobilized on western blot membranes. Advantages include (1) fast and easy; (2) purification of antibody with antigen in denatured form allows high yield in case antigen protein solubility is limited. Disadvantage is that possible antibodies that recognize certain 3D structure in solution of antigens might not be purified using such a method. Regarding this issue, the flow through is recommended to be kept and can be used for other purification methods with folded antigens., 该方案描述了从固定在western印迹膜上的变性抗原的血清中纯化抗体的方法。 优点包括（1）快速和容易; （2）用变性形式的抗原纯化抗体在抗原蛋白溶解度有限的情况下允许高产率。 缺点是在抗原溶液中识别某些3D结构的可能的抗体可能不能使用这种方法纯化。 关于这个问题，推荐保持流通，并且可以用于具有折叠抗原的其它纯化方法。