Multiparameter Flow Cytometry Analysis
Corresponding Organization : The Francis Crick Institute
Other organizations : Genomics England
Variable analysis
- Cell preparation method (peripheral blood, bone marrow, and spleen)
- Cell population analysis by flow cytometry
- Lysis of red blood cells in 155 mM NH4Cl, 12 mM NaHCO3, 0.1 mM EDTA buffer for 5 minutes at 4°C
- Washing of cells in PBS + 1% FBS before staining
- Lineage depletion of bone marrow and spleen cells using hematopoietic progenitor isolation kit
- Staining of cells with cell surface antibodies in PBS + 1% FBS for 1 hour at 4°C
- Washing of stained cells 3 times in PBS + 1% FBS
- Resuspension of stained cells in PBS + 1% FBS + 4',6-diamidino-2-phenylindole (0.1 μg/mL) before analysis
- Fixation of cells in 1.6% formaldehyde at room temperature for 10 minutes and permeabilization in Perm buffer III for intracellular staining
- Not explicitly mentioned
- Not explicitly mentioned
Annotations
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