Application: |
Flow Cytometry, intracellular staining |
Cells used: |
Mouse small intestinal lamina propria leukocytes (ILC2, CD4 T cells) |
Brief Protocol: |
Leukocytes were isolated from the small intestinal lamina propria. Cells were stimulated for 4 hours at 37oC with ionomycin (500ng/ml), phorbol 12,13-dibutyrate (500ng/ml) and brefeldin A (1000ng/ml). Cells were stained for surface markers (CD4, TCR, CD45) and viability, then fixed and permeabilized. Cytokine staining was performed overnight at 4oC in permeabilization buffer with antibodies at 600ng/ml. |
Results Summary: |
Approximately 2 orders of magnitude shift in IL-4 signal observed in response to stimulation. |
Additional Notes: |
Specificity confirmed using cells from IL-4-deficient mice. |
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