Hi Pavla,

attached is our protocol for PROTEIN-RNA CO-IMMUNOPRECIPITATION.

We have always been successful using Trizol RNA isolation following the IP.

We don't treat the samples with an RNAse inhibitor prior to the IP, but we apply a DNAse treatment to the IP prior to the Trizol extraction.  

In the protocol, you have the consideration for positive or negative controls.

Let me know if you have any questions and good luck,

Nicolas

Nicolas, thank you very much! That's exactly what I've been looking for. I will try it as soon as possible.

Pavla

Hi Nicolas,

Thanks for sharing the protocol. I am currently working on a project that requires RNA isolation after Protein-RNA Co-immunoprecipitation and your protocol is really helpful.

But I have a question. Why are you adding sepahrose beads A/G to the cell lysate? Is there a specific advantage by adding lysate to the beads and preclearing the lysate?