BD Biosciences
BD OptiBuild™ BUV805 Hamster Anti-Mouse CD3e
741928
500A2
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In different healthy and pathological contexts, CD4 T cells differentiate into a range of specialized interleukin-secreting subsets. In mouse models of aging and autoimmune diseases we study the plasticity and effector functions of CD4 T helper cells. We used anti-CD3e to further gate on the CD4 and CD8 T cell populations in old C57Bl/6J mice.
Flow Cytometry
Splenocytes of wild-type 24 months-old C57Bl/6J 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 1/200, anti-CD8 FITC, anti-CD4 BUV495, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C.
Cells were fixed and permeabilized with a commercial kit to counterstain for cytokines.
UV laser on Symphony cytometer
The CD3e positive population could be identified but the staining was not so good after stimulation of the cells.
N/A
Good to increase panel size.
Needs to be carefully titrated, signal spreading in other channels. Not very good after stimulation, expensive.
Usable but not optimal.