Yeast protein prep
- Grow 3-5 ml of yeast culture for each sample.
- Collect sample into Eppendorf tubes, freeze with liquid nitrogen to stop protein expression.
- To prepare cell lysates, boil the sample for 5 minutes in hot SDS-PAGE sample buffer (50 mM Tris-HCl pH 7.5, 5% SDS, 5% glycerol, 50 mM DDT, 5mM EDTA, complete protease inhibitors).
- Spin down at max speed for 1 minute. Collect the supernatant.
- Quantify the total protein content in the supernatant by Bradford assay reagent (Bio-rad).