Khan Dilshad H, Davie James R
Manitoba Institute of Child Health, 715 McDermot Avenue, University of Manitoba, Winnipeg, MB R3E 3P4, Canada.
Biochem Cell Biol. 2014 Aug;92(4):317-9. doi: 10.1139/bcb-2014-0028. Epub 2014 May 22.
Ribonucleoprotein immunoprecipitation (RIP) is an antibody-based method to detect RNA-protein interactions in situ. In the assay, UV cross-linking is commonly used to preserve RNA-protein interactions for subsequent target identification. UV light is a zero-length cross linker and thus identifies proteins directly bound to RNAs. Here, we describe a dual cross-linking RIP method that involves sequential protein-protein cross-linking step with a protein-protein cross-linker, followed by protein-RNA fixation by UV irradiation. In this way, proteins that indirectly bound to RNA can be analyzed.
核糖核蛋白免疫沉淀（RIP）是一种基于抗体的原位检测RNA-蛋白质相互作用的方法。在该检测中，通常使用紫外线交联来保留RNA-蛋白质相互作用以便后续进行靶标鉴定。紫外线是一种零长度交联剂，因此可识别直接与RNA结合的蛋白质。在此，我们描述了一种双重交联RIP方法，该方法包括先用蛋白质-蛋白质交联剂进行连续的蛋白质-蛋白质交联步骤，然后通过紫外线照射进行蛋白质-RNA固定。通过这种方式，可以分析间接与RNA结合的蛋白质。
