Madara P J, Banghart L R, Jack L J, Neira L M, Mather I H
Department of Animal Sciences, University of Maryland, College Park 20742.
Anal Biochem. 1990 Jun;187(2):246-50. doi: 10.1016/0003-2697(90)90451-e.
A procedure for the preparation of affinity-purified antibody is described. Protein mixtures are separated by electrophoresis in sodium dodecyl sulfate (SDS)--polyacrylamide gels. Individual bands of protein are cut from the gel and fixed in situ with glutaraldehyde. The gel pieces are then homogenized and washed extensively with buffered solutions and chaotropic agents. The washed gels can then be used as immunoadsorbents to purify antibodies from crude antisera. This method should be especially useful for the preparation of small amounts of antibody to proteins that are difficult to purify by conventional means, that are available only in limited quantity, or that cannot be blotted to immunoadsorbents such as nitrocellulose or diazotized paper.
本文描述了一种制备亲和纯化抗体的方法。蛋白质混合物通过在十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶中进行电泳分离。从凝胶上切下各个蛋白质条带，并用戊二醛原位固定。然后将凝胶块匀浆，并用缓冲溶液和离液剂广泛洗涤。洗涤后的凝胶随后可用作免疫吸附剂，从粗抗血清中纯化抗体。该方法对于制备针对难以通过常规方法纯化、仅以有限量获得或无法印迹到免疫吸附剂（如硝酸纤维素或重氮化纸）上的蛋白质的少量抗体特别有用。
