Biolegend
APC/Cyanine7 anti-human/mouse/rat CD278 (ICOS) Antibody
313529
C398.4A
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Our research aims at understanding how some CD4 T cells become autoreactive in melanoma patients treated with check point inhibitors. CD4 T cells are usually identified as CD3+ CD4+ cells. In this experiment, we used anti ICOS antibodies to identify ICOS+ CD4 T cells.
Flow Cytometry
Unstimulated healthy donor's PBMCs
Surface staining with anti- ICOS APC Cy7 at 1/200 and CD4, anti CD3, etc... (20' at 4°C, in the dark).
none
Staining with L/D dye (20' at 4°C, in the dark).
Cells were fixed and permeabilized with a commercial kit to counterstain for cytokines.
Red laser on Symphony cytometer
Memory CD4+ T cells (CD45RO+) express higher amounts of ICOS than naive CD4+ T cells (CD45RA+).
The antibody is specific and reliable.
The staining is not very bright. A FMO is needed to set the positive gate
Usable, but need some controls