When one studies the protein protein interaction, why it is sometimes recommended to perform in vitro linked transcription translation,
when immunoprecipitation anyhow confirms your in vivo results. I have done this in my masters thesis but have been asked by many that what is the objective. I could only say that its one more confirmatory assay apart from IP and one can be ascertained of direct interaction of your proteins as this assay is cell free system assay. Can anyone give me a better explanation 😀
I would say it’s a control, like you said to confirm the process. Although in vivo from IP supports your process of the protein-protein interaction there could be other factors within the organism affecting this interaction. When you take away the organism and only involve the particular molecules you take away some of the variability and hence increase the validity of your results.