Cell Surface Immunofluorescence Staining Protocol
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Cell Staining Buffer [ Details:BioLegend Cat. #420201]
Red Cell Lysis Buffer [ Details: BioLegend Cat. #420301]
7-AAD Viability Staining Solution[ Details: BioLegend Cat. #420403]
TruStain FcX™[ Details: anti-CD16/32, BioLegend Cat. #101319]
4. Cell-Surface Staining with Antibody:
9) Wash 2X with at least 2 ml of Cell Staining Buffer by centrifugation at 350 x g for 5 minutes.
4) Incubate on ice for 3-5 minutes in the dark.
5) Analyze with a Flow Cytometer.
5. Immunofluorescent Staining of Whole Blood:
2) Incubate at room temperature for 15-20 minutes in the dark.
4) Centrifuge at 350 X g for 5 minutes, discard the supernatant.
5) Wash 1X with at least 2 ml of Cell Staining Buffer by centrifugation at 350 x g for 5 minutes.
8) Resuspend cells in 0.5 ml Cell Staining Buffer or 0.5 ml 2% paraformaldehyde-PBS fixation buffer.