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| This protocol is for concentrating proteins from a variety of aqueous solutions. It is possible to scale up the protocol 1.5-fold and still fit the volumes in a microcentrifuge tube. |
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1. Put 0.1 ml (or as much as possible) of an aqueous protein sample in a 1.5 ml microcentrifuge tube.
2. Add 0.4 ml Methanol, 0.1 ml Chloroform (CAUTION! see Hint #1) and 0.3 ml ddH2O (water is optional) and mix with several inversions.
3. Centrifuge at 12,000 X g for 1 min (i.e. full speed in a microcentrifuge). This will separate the aqueous and organic phases and the proteins should be precipitated at the interface.
4. Remove the upper phase leaving behind the protein interface.
5. Add 0.3 ml of Methanol to give one phase.
6. Centrifuge at 12,000 X g for 2 min (i.e. full speed in a microcentrifuge). The proteins should pellet to the bottom of the tube.
7. Remove the supernatant and dissolve the proteins in the desired solvent.
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This bioProtocol does not use any solutions
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Methanol
Chloroform
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1. CAUTION! This substance is a biohazard. Consult this agent's MSDS for proper handling instructions.
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