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Brilliant Violet 605™ anti-human CD152 (CTLA-4) Antibody
369610
BNI3
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Our research aims to understand how some CD4 T cells become autoreactive in melanoma patients treated with check point inhibitors. In this experiment, we tested the expression of the inhibitory molecule CTLA-4 in CD4 T cells with an anti-CTLA-4 BV605 antibody.
Flow Cytometry
Unstimulated and stimulated healthy donor's PBMCs
Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.
None
Surface staining with anti-CD4 PE dazzle594, CD3 BUV395, CD45RO BUV737 etc., 30' at 4°C, in the dark, in BD Horizon™ Brilliant Stain Buffer.
Cells were fixed and permeabilized with commercial kit (FoxpP3 transcription factor kit, eBioscience) for 30 min at 4°C with fixative, 15 min at 4°C with permeabilization buffer, and stained with anti-CTLA-4 BV605 at 1/100 in permeabilization buffer.
Violet laser on Symphony cytometer
We could measure CTLA-4 on CD4 T cells. The antibody is relatively easy to use, although BV510 antibodies or L/D aqua have high spillover in 605 channel.
N/A
Gives a good signal.
L/D aqua has high spillover in 605 channel.
Good to expand a panel. Reliable.