Hi, I have received a few samples in Trizol at -80C, and I want to do a protein extraction for it. Now, I know that extraction using Trizol does not give good results, so is it possible to remove Trizol from the samples somehow and suspend them in lysis/RIPA buffer? Thanks
yes it is possible. After the phase separation step three layers will be formed:
a red organic phase (containing protein), an interphase (containing DNA), and a colorless upper aqueous phase (containing RNA). After removing the aqueous phase and interphase carefully by removing suoernatant you can save the rest for the protein isolation. precipitate the proteins using 1.5ml isopropanol per trizol used for the sample preparation. Allow samples to stand for at least 10 minutes at room temperature. Centrifuge at 12,000 × g for 10 minutes at 2–8 °C.
Discard the supernatant and add RIPA buffer to the lysate along with the protein inhibitor. Incubate on ice for 30 min. centrifuge at 10000rpm for 10 min.
Transfer supernatant to new tube for further analysis. you can store the samples at -80 for long term storage.
It will work.
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