BD Biosciences
PE Mouse Anti-TCF-7/TCF-1
564217
S33-966 (RUO)
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In different healthy and pathological contexts, CD4 and CD8 T cells differentiate into a range of specialized effector and regulatory subsets. In mouse models of aging and autoimmune diseases we study the exhausted phenotype of CD4 and CD8 T cells. We identified PD-1+TCF1+ cells with an anti-TCF-1 PE antibody in pre-lupus NXBxW F1 mice.
Flow Cytometry
Splenocytes of a NZBxW mouse, incubated in complete culture medium at 37 °C with (Restimulated) or without (unstimulated) PMA/ionomycin for 4h and brefeldin A for the 3 last hours.
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-CD3 BUV395, anti-PD-1 APC, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C
Cells were fixed and permeabilized with commercial kit (TrueNuclear Biolegend) for 45 min at 4°C with fixative, 15 min at 4°C with permeabilization buffer, and stained for cytokines and anti-TCF-1 PE 1/50 in permeabilization buffer.
Yellow-green laser on Symphony cytometer
We could identify and quantify the PD-1+TCF-1+ exhausted cells among CD4 and CD8 T cells. Most TCF-1+ cells are PD-1- naïve cells
N/A
Reliable and easy to use.
None
Good option, works on most cytometers, since it is conjugated to PE. Gives a good separation between positive and negative cells.