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PE/Cyanine7 anti-mouse CD152 Antibody
106314
UC10-4B9
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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 phenotype of the CD4 T regulatory cells. We identified CTLA-4-secreting cells with an anti-CTLA-4 PE-Cy7 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, CD4 BUV495, 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 with anti-FOXP3 FITC and anti-CTLA-4 PE-Cy7 1/100 in permeabilization buffer
Yellow-green laser on Symphony cytometer
We could identify and quantify CTLA-4-secreting FOXP3+ T regulatory cells. PMA+Ionomycin increases the proportion of CTLA4+ cells.
N/A
Bright, gives good separation.
A FMO should be done to set the positive threshold.
Good option, reliable, works with nuclear staining.