User:NYOUSHA Yousefi/Notebook/Protein Engineering/2012/10/09

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Entry title Protein expression Objective The frozen cells, that were presented with Zn Hb and were allowed to replicate, were prepared for further protein expression studies Procedure 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.