Gerber:Resources

Protocols

• RIP Protocol (Alessia Galgano) [show][hide]

Immunopurification protocol for the pulldown of RNA-binding proteins ("RIP") and co-purification of (associated) RNA molecules from human cell extracts using antibodies against endogenous proteins. RNA prepared according to this protocol can be analyzed using microarrays or sequencing techniques, allowing the identification of the RNA targets of the RBP under study. This protocol has been optimized for the immunopurification of endogenous human PUM1 and PUM2 proteins from HeLa S3 cells.
Reference:

1. Galgano A, Forrer M, Jaskiewicz L, Kanitz A, Zavolan M, and Gerber AP. Comparative analysis of mRNA targets for human PUF-family proteins suggests extensive interaction with the miRNA regulatory system. PLoS One. 2008 Sep 8;3(9):e3164. DOI:10.1371/journal.pone.0003164 |