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Immunopurification protocol for the pulldown of '''endogenous''' RNA-binding proteins ("RIP") and co-purification of (associated) RNA molecules from '''human''' cell extracts. 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.
 
Immunopurification protocol for the pulldown of '''endogenous''' RNA-binding proteins ("RIP") and co-purification of (associated) RNA molecules from '''human''' cell extracts. 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.<br/>
 
This protocol has been  optimized for the immunopurification of endogenous human PUM1 and PUM2 proteins from HeLa S3 cells.<br/>
Protocol from '''Alessia Galgano'''.
+
Protocol by '''Alessia Galgano'''.<br/>
 
<u>Reference:</u>
 
<u>Reference:</u>
 
<biblio>
 
<biblio>

Revision as of 06:21, 26 September 2008

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Protocols

Immunopurification protocol for the pulldown of endogenous RNA-binding proteins ("RIP") and co-purification of (associated) RNA molecules from human cell extracts. 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.
Protocol by Alessia Galgano.
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 | PubMed ID:18776931 | HubMed [Paper17]