Trieu Edward P, Gross Joanne K, Targoff Ira N
Arthritis and immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104, USA.
Methods Mol Biol. 2009;536:259-75. doi: 10.1007/978-1-59745-542-8_28.
Combining the procedures of immunoprecipitation and immunoblotting can help to 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 to 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" immunoprecpitation-blotting), is reported here in detail.
将免疫沉淀和免疫印迹程序相结合有助于克服每个单独程序的一些局限性。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、更灵敏地识别免疫沉淀的蛋白质。与全细胞提取物的免疫印迹相比，免疫沉淀可以富集感兴趣的蛋白质，以提高检测灵敏度。最近，免疫沉淀-印迹有助于鉴定一种新的自身抗体——抗-p155，并检测患者血清中该自身抗体的存在，以研究其临床关联。本文详细报道了免疫沉淀-印迹程序，特别是针对这种自身抗体检测（“反向”免疫沉淀-印迹）。
