Cell Fractionation

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Cell Fractionation
This separates different parts and organelles of a cell so study can be
undertaken in detail.
Stages
1. Cut tissue and place in a cold isotonic buffer. Cold to stop enzyme
action, isotonic to stop osmosis and buffer to stop changes in PH. Grind to
break open cells.
2. Filter, removes insoluble tissue, e.g. fat connective tissue, cell walls etc.
3. Centrifuge at low speed: 1000x g for 10 mins: this pellets nuclei.
4. Centrifuge at medium speed: 10000 x g for 30 mins: this pellets
mitochondria and chloroplasts.
5. Centrifuge at high speed 100000 x g for an hour: this pellets ER, Golgi
and other membrane fragments.
6. Centrifuge at very high speed 300000 x g for 3 hours: pellets ribosomes.
7. Now organelle free cytoplasm.

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