I am planning to make small RNA molecules and cap them. I want to check the ability of these m7G RNA's to eIF4E/eIF4F complex, whichever may be easier. Has anyone done anything like this before and is it an easy technique. I am new to this kind of assay. Any help would be much appreciated.
You have at least two different approaches: 1) RIP (RNA immuno-precipitation) which requires an antibody against the protein of interest. You pull down the protein and do a RT-PCR. 2) RNA-pull down, which requires in-vitro transcription of your RNA and adding a biotin molecule to it. Then a pull-down by Strep-beads followed by western blot.
Thanks for both the approaches. Is there a follow up protocol that you have used or know that works well. Also would you need to add PABP to translate?
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