Application: |
Flow Cytometry |
Cells used: |
Cell suspensions from mouse lymphoid tissues such as spleen and lymph nodes, (but also peripheral non-lymphoid tissues such as brain, lung, liver, and tumour). |
Brief Protocol: |
We isolated cells from tumour implanted in from C57BL/6 mice, by using collagenase/ DNase treatment for 30min at 37C in a shaker. We put up to 5 x 10^6 cells in a 50ul volume and add BV711 anti-mouse CD8a antibody diluted at 1/300 final in FACS buffer for 1h at room temperature. |
Results Summary: |
Anti-mouse CD8a antibody coupled to BV711provides a reasonable signal that allows reliable staining to study CD8 T lymphocytes from various types of mouse tissues, including non-lymphoid organs that require a collagenase/ DNase treatment. |
Additional Notes: |
Our laboratory has a long lasting interest in immune responses and we have tested many different fluorescent antibodies to detect lymphocyte phenotype, activation and effector functions. We use routinely BV711 anti-mouse CD8a antibody in our combination with more 9 other colours. Figure shows results of tumour infiltrating live CD3+ CD45+ cells stained with BV711 anti-mouse CD8a antibody and BV605 anti-mouse CD4 antibody. |
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