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Standard ProtocolsIsolation of complexes from serum | ||
Complexes can be isolated from serum by centrifugation on either a sucrose or Percoll gradient. The sucrose gradient is the easier of the two to set up.
Sucrose gradient:
Isolating complexes from a Percoll gradient requires that an optimal gradient be determined for each complex to be isolated. For example, pLL/DNA complexes (pLL Mw 24,000) centrifuge into a Percoll layer of 1.04 g/ml. The best separation of serum from complexes was found using a gradient consisting of 2 mls of water, 3 mls 1.01 g/ml Percoll, 3 mls 1.02 g/ml, 2 mls 1.03 g/ml, 1 ml 1.04 g/ml, 1 ml 1.06 g/ml and 1 ml of 1.1 g/ml. Other complexes required different gradients for optimal separation of complexes from serum. |
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