Protein extraction from mouse soft tissues
Instrument: Minilys homogenizer
Sample: Mouse spleen, colon, lung
Protocol:
- Precellys kit: 03961-1-004 (VK05 glass beads).
- Add ~50 mg of organ samples (spleen, colon, lung) flash-frozen in liquid nitrogen and stored at -20°C. Thawed on ice prior to lysis.
- Add Buffer: 250μl of RIPA buffer supplemented with 10% Protease inhibitor (Sigma P8340).
- Run the Minilys at 5000 rpm, 3x60sec (in ice between each cycle).
- After lysis, centrifuge tubes at 10000 rpm for 10 min. Obtain supernatant for the further analysis.
Results:
The protein content of the samples was estimated with the QuiantiT Protein assay kit Invitrogen Q33211, using a 100-fold dilution of the original sample. Excellent results were obtained from different mice tissues using the Minilys apparatus for tissue lysis.
Protein recovery in 100μl buffer volume compared to the initial weight of the lysed spleen and colon tissue obtained with Minilys.
