TCA precipitation
Make 100% TCA, 3g in 3mls of filtered water, ice it
Mix SILAC samples: L, M, H together in equal volumes or equal protein amount
Add TCA to a final concentration of 10%, i.e.: 60ul of TCA in 600ul sample
Incubate in ice 10 min
Centrifuge at 4 C at max speed 10 min
Wash with 500 ul cold acetone (store acetone at -20 C)
Vortex 2X ~10 sec
Incubate at -20 C 5min
Spin 15 max speed at 4 C
Wash again with 500ul cold acetone
Vortex lightly
Spin 15 min at 4 C
Remove acetone
Evaporate in hood 1-2 min (don’t dry pellet completely)