Hi,
I am looking to study a membrane protein in N.tabacum and I need to find a way to separate the protein from the membrane in order to extract it using Immunoprecipitation.
I know that lysis would have to be involved but am finding it hard to locate a protocol that allows me to do it in plants, specifically N.tabacum and would like to know if anyone has any experience in doing so.
Thanks,
Barry
Sorry, I meant to add that the protein I was looking at was in the outer Mitochondrial membrane.
Dear Barry,
i think the best is to first purify Mitochondria and then extract your target protein form the outer membrane. I suggest you consider using BY-2 cells as starting material because the cell walls are more easily digested compared to leaf-tissue. You may even look for or make a cell line that has a GFP or DsRed targeted to mitochondria or the outer membrane to help you identify the fraction you are interested in.
Good luck! Markus
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