Trieu Edward P, Targoff Ira N
Arthritis and Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA.
Methods Mol Biol. 2015;1312:327-42. doi: 10.1007/978-1-4939-2694-7_34.
Combining the procedures of immunoprecipitation and immunoblotting can help overcome some of the limitations of each separate procedure. Immunoblotting can identify immunoprecipitated proteins more specifically and with higher sensitivity than nonspecific protein stains or autoradiography. Immunoprecipitation can enrich proteins of interest to improve sensitivity for detection when compared with immunoblotting of whole cell extracts. Recently, immunoprecipitation-blotting helped us characterize a new autoantibody, anti-p155, and to test for the presence of the autoantibody in patient sera to study its clinical associations. The procedure for immunoprecipitation-blotting, with specific reference to this autoantibody test ("reverse" immunoprecipitation-blotting), is reported here in detail.
将免疫沉淀和免疫印迹这两种方法结合起来，有助于克服每种单独方法的一些局限性。与非特异性蛋白质染色或放射自显影相比，免疫印迹能够更特异性、更灵敏地鉴定免疫沉淀的蛋白质。与全细胞提取物的免疫印迹相比，免疫沉淀能够富集目标蛋白，从而提高检测的灵敏度。最近，免疫沉淀-印迹法帮助我们鉴定了一种新的自身抗体——抗-p155，并检测患者血清中该自身抗体的存在情况，以研究其临床关联。本文详细报道了免疫沉淀-印迹法的操作步骤，特别针对这种自身抗体检测（“反向”免疫沉淀-印迹法）。
