Protein extraction from Arabidopsis with Minilys
Instrument: Minilys homogenizer
Sample: Arabidopsis seedlings
- Precellys kit: 03961-1-010 (1.4 & 2.8 ceramic beads mix).
- Add samples 5 Arabidopsis seedlings (two weeks old).
- Add buffer 200 μL of 62.5mM Tris-HCl pH8, 4M Urea, 3% SDS, 10% Glycerol, 0.1% Bromophenolblue, 100mM DTT.
- Snap-frozen in N2.
- Addition of extraction buffer after (Preparation A) or before homogenization (Preparation B).
- Run the Minilys at 5000 rpm, 20 sec or 5000 rpm, 40s.
- Triplicate by condition.
Total proteins were separated by western blot on a 8% SDSAcrylamide gel and transfer to a PVDF-membrane.
Western blot of Arabidopsis total proteins and detection of HD-Zip transcription factor by anti GFPHRP antibody.