1- Grow approximately 10,000 worms on 2 x 150 mm NGM plates seeded with E. coli OP50 bacteria.
2- Harvest worms and wash them 3 times with 15 ml of M9 buffer.
3- Wash worms once with 15 ml of ddH2O.
4- Resuspend worms in 3 ml of ice cold MIB and transfer them into a 7 ml dounce homogenizer on ice.
5- Perform 21 gentle strokes.
6- Centrifuge at 200 g for 5 min and keep supernatant.
7- Centrifuge at 800 g for 10 min and keep supernatant.
8- Centrifuge at high-speed 12,000 g for 10 min to pellet the total mitochondria.
9- MIB is removed and replaced by 500 µl of ice cold PEB.
10- Add 100 µg of anti-HA magnetic beads.
11- Incubate samples at 4 °C for 1 h while rotating at 10 rpm.
12- Transfer the samples on a homemade setup consisting of a column and a magnet.
13- Ensure that beads were attracted to the side of the tip and held by the magnet and then wash twice with 25 ml of ice cold PEB.
14- After washings, resuspend the beads in 20 µl of MIB.