Tumor dissociation for flow cytometry analysis
Digest medium: collagenase A (1mg/ml,Roche 10103578001), DNAseI (105 units/ml,Roche 04716728001) in RPMI
RBS lysis buffer
1. Dissect tumor and dice with razor.
2. Digest tumor in digest medium for 30 min-1hr (2ml digestion medium/tumor). Quench digestion with 0.5ml FBS and pass through strainers.
3. Wash with PBS1x.
4. Incubate with 5ml RBC lysis buffer for 5mins.
6. Quench with 45ml PBS. Spin down.