Biolegend
Alexa Fluor® 647 anti-TCF1 (TCF7) Antibody
655204
7F11A10
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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-TCF-1 Al647 antibody to identify TCF1+ CD4 T cells.
Flow Cytometry
PMA+ionomycin+Calcium-activated healthy donor's PBMCs
Surface staining with anti-CD3, anti-CD4 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 cytoplasmic proteins, including cytokines. The anti-TCF1 Al647 was added on the cells at 1/50 in perm/wash buffer and incubated for 30' at 4°C, in the dark.
Red laser on Symphony cytometer
Cytoplasmic TCF-1 can be seen on CD4+ CD3+ T cells even without nuclear permeabilization.
The antibody is specific.
The antibody gives good signal, but a FMO control is important to set the positive threshold.
Can be used