Yeast Crude Protein Extraction
Grow cells to mid - exponential phase in liquid medium .
Normalize cells to an OD600 of 2 - 3 in 1 mL of H2O .
Centrifuge 1 mL of H2O containing your cells at 9000 RPM for two minutes .
Resuspend pelleted cells in 100 uL 0.2 M NaOH and incubate on ice for 15 minutes .
Centrifuge cells at 9000 RPM for two minutes .
Resuspend the pellet in 100 uL of Yeast Protein Extraction Sample Buffer .
Add 0.4 uL 2 - mercaptoethanol to the mixture .
Boil cells in sample buffer for 5 minutes at 95 C .
Can store at - 80 C or load supernatant into a gel .
Vortex briefly then centrifuge at 10,000 RPM for 3 minutes .
