BD Biosciences
Fixation/Permeabilization Solution Kit
554714
Splenocytes from mice were stimulated for 6 hrs with PMA and Ionomycin in the presence of protein transport inhibitor Golgi Stop. Cells were harvested and surface stained with PE conjugated rat anti-mouse CD8 antibody. These cells were later fixed and permeabilized using a Fixation/Permeabilization Solution Kit followed by staining with rat anti-mouse IFN-gamma antibody.
Flow cytometry
Mouse splenocytes
1. Stimulated cells were first fixed for 15 minutes with the cytofix buffer at room temperature 2. These fixed cells were washed twice with 1X perm wash buffrer 3. Fixed and permeabiized cells were stained with IFN-gamma specific antibody for 30 min. 4. Cells were washed and analyzed by flow cytometry.
Never mix with pipettes and gentle mixing is enough
Presence of IFN-gamma positive cells confirm the usefulness of this kit.
None
Very good kit for intracellular staining
Very useful for intracellular staining