Nuclear Factor Fixation and Permeabilization Staining Protocol
Prepare target cells of interest and perform surface staining as described in BioLegend's Cell Surface Immunofluorescence Staining Protocol .
Fix the cells with 1 ml / tube BioLegend's Nuclear Factor Fixation Buffer , at room temperature in the dark for 20 minutes .
Centrifuge at 250 x g for 5 minutes ; discard supernatant .
Wash once with 1 ml Biolegend's Nuclear Factor Permeabilization Buffer ( 1x ) .
Resuspend cells in 1 ml BioLegend's Nuclear Factor Permabilization Buffer ( 1x ) , incubate at room temperature in the dark for 20 minutes , spin down cells and discard the supernatant ; then resuspend the pellet in 100 ul of BioLegend's Nucelar Factor Permeabilization Buffer ( 1x ) .
Add appropriate amount of flurochrome - conjugated antibodies for nuclear target of interest , and incubate at room temperature in the dark for 30 minutes .
Wash twice with Cell Staining Buffer , and resuspend in 0.5 ml Cell Staining Buffer , then analyze with flow cytometer with appropriate instrument settings .
