[PDF] cell fixation protocol for flow cytometry

What are the flow cytometry protocols?

The flow cytometry protocols below provide detailed procedures for the treatment and staining of cells prior to using a flow cytometer. Direct staining of cells applicable where the fluorophore is directly linked to the primary antibody

How do I use a fixation and permeabilization kit for flow cytometry?

Alternatively, you can use our fixation and permeabilization kit for flow cytometry ( ab185917 ), which is suitable for most sample types. Fix the cells in your chosen fixative. Spin down cells to a pellet (200 g, 5 mins, 4°C), discard the supernatant, and resuspend the pellet in fixative. Incubate cells with the fixative as indicated below.

How do you use a flow cytometry staining buffer?

Combine the recommended quantity of each primary or fluorophore-labeled antibody in an appropriate volume of Flow Cytometry Staining Buffer so that the final staining volume is 100 µL (i.e., 50 µL of cell sample + 50 µL of antibody mix) and add to cells. Pulse vortex gently to mix. Incubate for 30 minutes at 2–8°C. Protect from light.

How do you fix cells with a fixative?

Fix the cells in your chosen fixative. Spin down cells to a pellet (200 g, 5 mins, 4°C), discard the supernatant, and resuspend the pellet in fixative. Incubate cells with the fixative as indicated below. Tip: The fixation will require optimization for different antigens.