Preparation of Intracellular Fraction

1. Add 1-2 g of glass beads to the resuspended cells.

2. Disrupt the cells five times for 1 min under CO2 atmosphere.

3. Cool the samples on ice for 5 min between each mixing sequence.

4. Centrifuge the sample at 20,000g for 30 min at 4°C to remove glass beads and unbroken cells (see Note 2).

5. Transfer the supernatant (intracellular fraction) to a clean tube by carefully decanting from the pellet and maintain on ice (see Note 3).

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