Total protein extraction kit buffer

Fly heads were homogenized using an ultrasonicator at 4 °C for 6 min at 40 kHz in 3 µL of total protein extraction kit TM buffer containing 0.1% SDS (Millipore). The homogenized lysates were centrifuged at 17,000 × g for 1 min at RT to remove the debris. The supernatant was collected and centrifuged again under the same conditions to collect the fresh supernatant. Four-week-old mnd2/mnd2 mice, mnd2/+ mice and aged match controls were sacrificed, and the brain tissues were chopped into small pieces, followed by the addition of 1 mL of total protein extraction kit TM buffer containing 0.1% SDS (Millipore) to 0.2 g of brain samples. Samples were homogenized, and total proteins were isolated as described above.