Tumor and Immune Cell Analysis Protocol
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Corresponding Organization :
Other organizations : Memorial Sloan Kettering Cancer Center
Protocol cited in 1 other protocol
Variable analysis
- Tumor mincing
- Incubation in 12 mg/mL type 2 collagenase (Worthington Biochemical) plus 0.5 mg/mL DNAse I (Roche Diagnostics) for 30 minutes at 37°C
- Mechanical dissociation of tumors and draining lymph nodes (DLN)
- Mashing of spleens through a 70 μm strainer
- Incubation in ammonium chloride lysis buffer (eBioscience)
- Flushing of bone marrow from one tibia per mouse
- KIT+ cell analysis
- Macrophage analysis
- T cell analysis
- Washing through 100 then 40 μm nylon cell strainers (Falcon, BD Biosciences) with 1% fetal calf serum (FCS)
- Quenching of ammonium chloride lysis buffer in 1% FCS
- Washing of cells in 1% FCS
- Not explicitly mentioned
- Not explicitly mentioned
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