I am interested in making a Chlamy cell lysate for use in
pulling-down binding partners to a GST-fusion protein expressed
in bacteria and then bound to glutathione beads.
Thus, I would like to be able to get at the cytosolic proteins
leaving behind the nuclei, chloroplasts, and the associated DNA.
Does anyone have a protocol for doing this?
I suppose it would start with getting rid of the cell wall,
followed by disruption either mechanically and/or by mild
detergents and finally ending with pelleting of insoluble
Thank you very much.
Joshua Z. Rappoport PhD
The Rockefeller University
Laboratory of Cellular Biophysics
1230 York Avenue, Box 304
New York, NY 10021