Miltenyi
CD45RO Antibody, anti-human, VioBlue®, REAfinity™
130-119-710
REA611
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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 CD45RO VioBlue antibody to identify memory CD4 and CD8 T cells.
Flow Cytometry
PMA+ionomycin+Calcium-activated healthy donor's PBMCs
Surface staining with anti-CD45RO VioBlue at 1/100 (see titration), anti-CD45RA, anti-CD3, anti-CD4, anti CD8, 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.
Violet laser on Symphony cytometer
CD45RO can be seen on CD4+ CD3+ T cells but the antibody should be carefully titrated, as it does not work well if too concentrated, and the signal is rapidly lost above 1/100 dilution.
The antibody is specific.
This antibody needs careful titration and is not very bright.
Can be used, , but not ideal on activated cells