Maheshri:ProteinPrepY

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Yeast protein prep

1. Grow 3-5 ml of yeast culture for each sample.
2. Collect sample into Eppendorf tubes, freeze with liquid nitrogen to stop protein expression.
3. 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).
4. Spin down at max speed for 1 minute. Collect the supernatant.
5. Quantify the total protein content in the supernatant by Bradford assay reagent (Bio-rad).