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Example of immunofluorescent staining with the Treg Detection Kit (CD4/CD25/CD127) using whole blood and the lyse/no wash protocol with CD45 trigger. A–E depict the gating strategy for the flow cytometric analysis of CD4+
An appropriate threshold was defined based on CD45-VioBlue versus side scatter signals, for the exclusion of debris and erythrocytes from the data.
Lymphocytes were gated for their expected morphology using a forward scatter versus side scatter "lymphocyte gate" (P1)
Dead cells were excluded by a CD127-PE versus propidium iodide dot plot (P2).
lymphocytes were gated in P3.
Treg cells were gated by a CD127-PE versus CD25-APC dot plot.
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