Wiser M F, Schweiger H G
Anal Biochem. 1986 May 15;155(1):71-7. doi: 10.1016/0003-2697(86)90226-5.
The sensitivity in antigen detection during immunoblot analysis is greatly increased if the antigen is first immunoprecipitated from the crude extract before electrophoresis and transfer to nitrocellulose. Not only does the method allow detection of antigens which are minor components of crude mixtures or antigens which cannot be radiolabeled, but the method also resolves problems, such as high background, which are often associated with immunoprecipitation. Also, by modifying the method, whether or not monoclonal antibodies recognize the same or different antigens and/or epitopes can be easily determined.
如果在电泳和转印至硝酸纤维素膜之前,先从粗提物中免疫沉淀抗原,那么免疫印迹分析中抗原检测的灵敏度会大大提高。该方法不仅能够检测粗混合物中的次要成分抗原或无法进行放射性标记的抗原,还能解决诸如高背景等常常与免疫沉淀相关的问题。此外,通过改进该方法,可以轻松确定单克隆抗体是否识别相同或不同的抗原和/或表位。