Wittrup:Tumor dissociation

Tumor dissociation for flow cytometry analysis

Material:

Digest medium: collagenase A (1mg/ml), DNAseI (105 units/ml) 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.