If you are running controls at FACS:
The amount of Blocking Buffer we will need to resuspend our nuclei pellet with after centrifuging depends on the number of controls we will run. Each control tube will require 50 μL of nuclei suspension, and our sample needs to have 1,000 μL of nuclei suspension left over. In this case we are running two controls.
After the homogenate is done centrifuging, discard the supernatant and resuspend the pellet in 1,100 μL of Blocking Buffer. Incubate for 10 minutes on ice. Add 950 μL Blocking Buffer to each of your control tubes. Add 50 μL of nuclei suspension to each control tube.
Add the corresponding antibodies to the control tubes:
Place sample and controls in a rotator placed in the dark at 4°C and incubate for 30 minutes.