Protein extraction from Arabidopsis with Minilys

Instrument: Minilys homogenizer

Sample: Arabidopsis seedlings

Protocol:

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.

Results:

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.