Wittrup:Tumor dissociation
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Tumor dissociation for flow cytometry analysis
Material:
Digest medium: collagenase A (1mg/ml,Roche 10103578001), DNAseI (105 units/ml,Roche 04716728001) in RPMI
RBS lysis buffer
Strainer
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.
7. Stain.
