Cross Linking and Immunoprecipitation (CLIP)
(Laboratory of Molecular Neuro-Oncology, Rochefeller Univ.)
CLIP or cross-linking and immunoprecipitation assay exploits the covalent protein-nucleic acid cross-linking to stringently purify a specific protein-RNA complex using immunoprecipitation followed by SDS-PAGE separation. In this way, the vast majority of non-specific contaminating RNA, which can bind to co-immunoprecipitated proteins or beads, can be removed. Here is an improved protocol that performs RNA linker ligation before the SDS-PAGE step, and describe its application to the specific purification and amplification of RNA ligands of Nova in neurons.
Added: Sat Jan 31 2009, Hits: 4920, Reviews: 0