Application: |
Flow cytometry |
Cells used: |
Splenocytes from naive C57Bl/6 mouse |
Brief Protocol: |
Three millions splenocytes prepared in FACS buffer (PBS/Bovine Serum Albumin 1% / 0.1% azide) were plated in round-bottom 96-well plates. Cells were spin down and treated with Fc block (5% of supernatant from a 2.4 G2 hybridoma culture in FACS buffer), for 10 min at 4°C. Cells were spin down and stained in 25 µl of FACS buffer with BV421™ anti-CD11b (1:1300) during 20 min at 4°C in the dark. Cells were washed twice with 150 µl of FACS buffer PBS/BSA, and resuspended in FACS buffer for analysis by flow cytometry. |
Results Summary: |
Identification of MHC class II negative granulocytes and monocyte/macrophages and a subset of MHC Class II positive dendritic cells among TCRbeta negative/IgM negative gate. |
Additional Notes: |
Identification of myeloid cells and B1 cells by flow cytometry. |
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