TCA Precipitation of Proteins
Preparation of 100% TCA: (don't try to weigh out TCA; it's too hygroscopic)
- Obtain a fresh bottle of crystalline TCA.
- Read the weight in the container from the label.
- Add distilled water to give a 100 g / 100 ml solution at final volume.
0.01 M HCl/90% acetone
- Add 100% (w/v) TCA (trichloroacetic acid, ( see preparation method above) to the sample to bring the TCA concentration to 20%.
- Incubate on ice for at least 1 hr. Diluted samples may be left overnight.
- Spin at maximum speed in a microcentrifuge for 10 min.
- Wash the pellet 3X with a solution of ice cold 0.01 M HCl / 90% acetone. Allow the pellet to air dry.
The pellet can then be resuspended directly in 100 mM Tris, pH 8.5, 8 M urea or 6 M Guanidine- HCl in 25 mM ammonium bicarbonate buffer for enzymatic digestion for mass spectrometry
Last modified: Jul 26, 2018