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[Bio101] Protein Precipitation from Detergent-containing Samples   

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In this protocol

Abstract

Materials and Reagents

  1. Methanol
  2. Chloroform

Equipment

  1. Centrifuge

Procedure

For 100 μl of sample:

  1. Add 300 μl of methanol to the protein sample and mix.
  2. Add 100 μl of chloroform and mix. Mixing yields a single phase.
  3. Add 200 μl of water and mix vigorously. Initiation of phase separation.
  4. Centrifuge at 10,000 x g for 2 min. Precipitated proteins are at the interface.
  5. Remove upper aqueous methanol phase.
  6. Add 300 μl of methanol to the bottom phase and mix. Mixing yields a single phase.
  7. Centrifuge at 10,000 x g for 1 min.
  8. Air-dry the protein pellet with the tube inverted.

References

  1. Whitelegge J.P., le Coutre J., Lee J.C., Engel C.K., Prive G.G., Faull K.F., Kaback H.R. (1999). Toward the bilayer proteome, electrospray ionization-mass spectrometry of large, intact transmembrane proteins. Proceedings of the National Academy of Sciences of the United States of America 96(19): 10695-8.
Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Aboulaich, N. (2011). Protein Precipitation from Detergent-containing Samples. Bio-protocol Bio101: e40. DOI: 10.21769/BioProtoc.40.
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