Protocol 1. Pellet induced bacterial cells by centrifugation at 1,000 rpms for 5 minutes. Add the bacterial into PBS, suspense and centrifugate to discard PBS. 2. Add the precooling lysate solution (ml) to the bacterial cell (g) in 10-50:1 (i.e. Add 10-50ml lysate solution to 1g bacterial cell) to lyse and shake for 30-60 min. 3. If there's an ultrasonator, lash the cells uder the maximal frequency shortly for 4 times, 10-30s each. (spurging is not the standard for the power), during the ultrasonic interval, put the cells into the ice water to reduce the temperature. 4. Centrifuge at 10,000 g at 4°C for 10 minutes. 5. Blot the supernatant into a centrifuge tube carefully and place it on the ice. (Note: If necessary, centrifuge at 10,000 g for 30 minutes at 4°C the remove the agglomerative protein. Then blot the supernatant.) 6. The supernatant could be used for albuminimetry.Experiment Notes: Lyses bacterial and cell gentle, can extract recombination protein efficiently. Easy and quick operation, and the lysate solution can be dialyzed.
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