Lee Choogon
Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, USA.
Methods Mol Biol. 2007;362:401-6. doi: 10.1007/978-1-59745-257-1_31.
As with most other proteins, clock proteins physically interact with one another. Coimmunoprecipitation (coIP) is the most straightforward technique to study protein-protein interactions in vivo, if antibodies against the proteins of interest are available. To perform coIP, first an antibody against a target protein is coupled to Sepharose beads through protein A or G, then the complexes containing the target protein are immunoprecipitated with the antibody-coupled beads by centrifugation. Protein components in the complexes are visualized by Western blotting using antibodies specific to the different components.
与大多数其他蛋白质一样，生物钟蛋白之间会发生物理相互作用。如果有针对目标蛋白质的抗体，免疫共沉淀（coIP）是研究体内蛋白质 - 蛋白质相互作用最直接的技术。进行免疫共沉淀时，首先将针对目标蛋白的抗体通过蛋白A或G与琼脂糖珠偶联，然后通过离心用抗体偶联的珠子免疫沉淀含有目标蛋白的复合物。复合物中的蛋白质成分通过使用针对不同成分的特异性抗体进行蛋白质印迹法来可视化。
