The frozen cells, that were presented with Zn Hb and were allowed to replicate, were prepared for further protein expression studies
- The zinc protoporphyrin prepared in previous labs was presented to cultured cells.
- The containers of the frozen cells were taken out of the freezer and were allowed to slowly cool down at the room temperature in a beaker full of tap water.
- When the cells mixture completely turned into liquid, the cells were killed and broken apart by a ultrasound processor in 30seconds intervals of ultrasound and 30s intervals of sitting still in the ice. This intervals were repeated for 3 times.
- The cell mixtures were placed into special centrifuge tubes and their weights were balanced to three decimal places.
- The cells were centrifuges at 18000g for two hours.
- After the centrifuge, the supernatant was separated and stored in different tubes.