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8.2.4.1.1 Centrifugation
/span>The enzyme extract is centrifuged to remove cell debris, cell organelles and sometimes other molecular aggregates, leading to partial purification of enzymes. It also helps in characterization of an enzyme, since, depending upon its mass and shape the enzyme will move through a solution at a definite speed and occupy a characteristic position in the centrifuge tube. For most cytosolic enzymes, centrifugation at about 30,000 g for 30 minutes is good enough to obtain a fair amount of activity in the supernatant. However, if the enzyme is located in a specific cell organelle, an extract rich in that organelle is prepared through 'preparative centrifugation'. (Centrifugation for different durations at different velocities allows the cell organelles to sediment according to their sizes). All centrifugation operations are conducted in cold(O-4°C). |
Last modified: Saturday, 12 November 2011, 6:45 AM