BioLegend
Pacific Blue™ anti-mouse/human Helios Antibody
137220
22F6
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In different healthy and pathological contexts, CD4 T cells differentiate into a range of specialized subsets, including FOXP3+ regulatory T cells expressing different levels of CD25 and transcription factor Helios. In mouse models of aging and autoimmune diseases we study the plasticity of effector and regulatory CD4 T cells. We used anti-Helios Pacific Blue to analyze the CD4+FOXP3+ T cell population in healthy control BALB/C mice.
Flow Cytometry
Splenocytes of wild-type 6 months-old BALB/C mouse
Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.
2.4G2
Surface staining with anti-CD3 BUV805, anti-CD4 BUV495, anti-CD25 PerCP Cy5.5, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C.
Cells are then fixed and permeabilized with commercial kit (True Nuclear, BioLegend) 45 min at 4°C with fixative, and 15 min at 4°C with permeabilization buffer) and stained with an anti-FOXP3 FITC and the anti-Helios Pacific Blue at 1/50.
Violet laser on Symphony cytometer
The staining identifies different FOXP3+ subpopulation that express different levels of CD25 and Helios.
N/A
Bright and gives good resolution.
None
Very good options, easy to use.