Hi Steemit scientists and enthusiasts,
I wanted to share an obscure method of the protein purification process. This it might be the last time I use this machine before I graduate.
The French pressure cell press was most commonly used in the 70's, and has since been replaced with less bulky equipment; however, the efficiency of the French press is hard to compete with. I dont think many, if any, are currently in production.
In order study the biochemistry of a protein, the protein of interest needs to be synthesized, and then purified. This post will be briefly covering one step of the protein purification process - preparation of a cell lysate by French press.
First, we cultivate a lot of E.coli cells that have been instructed to synthesize our protein of interest (A topic for another blog). The cells are put into the pressure cell which can handle 20,000 PSI...
A hydraulic press forces the E.coli cells through an aperture that is smaller than the cell can pass through whole. The cell membranes are disrupted, and all of the cytosolic proteins are collected. The cell lysate is then put into a centerfuge, and spun at 14,000 RPM to separate the cell membranes from the soluble protein.
The next step in the protein purification process is to separate the protein of interest from the other endogenous E.coli proteins with column chromatography a Nickel-NTA resin. I will cover in another post...
Thanks for checking this out. If you are a fellow scientist, let me know how you lyse cells in the comments...
-The Scientist