Protein extraction from arabidopis with minilys - Monitoring and Testing - Laboratory Equipment
- Minilys homogenizer.
- Precellys kit: 03961-1-010 (1.4&2.8 ceramic beads mix).
- Samples: 5 Arabidopsis seedlings (two weeks old).
- 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).
- Minilys: 5000 rpm, 20 sec vs 5000 rpm, 40s. -Triplicate by condition.
- Analysis: Western blot.
The total yield of proteins extracted is increasing with buffer extraction and an higher duration of homogenization. Total proteins were separated on a 8% SDSAcrylamide gel and transfer to a PVDF-membrane.
Buffer extraction with an increasing time homogenization allow higher proteins yield and in the same range as with Precellys 24 tissue homogenizer. Minilys provides the optimal balance of efficiency, speed, ease of use with a low throughput. Minilys enables cross-contamination free homogenization as opposed grinding with a mortar.