TCA protein precipitation

Kyle Coachman/Linda Warfield/Steve Hahn

Last modified 5/13/02

To concentrate proteins for analysis by SDS PAGE:

If a small amount of protein is to be precipitated (less than a few micrograms), add Insulin as a carrier protein (10 micrograms of Sigma insulin, I-5500, per sample works well).

  1. Add an equal volume of 20% TCA (trichloroacetic acid) to protein sample.
  2. Incubate 30 min on ice.
  3. Spin in microfuge at 4 deg. For 15 min.
  4. Carefully remove all supernatant.
  5. Add ~300 ul cold acetone and spin 5 min at 4 degrees.
  6. Remove supernatant and dry pellet.
  7. Resuspend samples in SDS PAGE loading buffer. Load to SDS PAGE after heating at 65 deg for 3 min.
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