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Brilliant Violet 650 anti-mouse CXCR5 Antibody
145517
L138D7
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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-CXCR5 to quantify the number of PD1high CXCR5+ CD4 T follicular helper cells in old C57Bl/6J mice and study their cytokine production.
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.
Surface staining with BV650 anti-CXCR5 at 1/300, PerCP efluor710 anti-PD1 antibody, anti-CD3 Alexa700 and anti-CD4 Pacific Blue in PBS 0.5% BSA incubated 30min at 4°C. Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.
Clone 2.4G2 (Fc block)
Cells are then fixed and permeabilized with commercial kit (30 min at 4°C with fixative, and 15 min at 4°C with permeabilization buffer). Cells are finally incubated with anti-cytokine antibodies in permeabilization buffer for 30 min at 4°C and washed once with permeabilization buffer and once with PBS BSA 0.5%.
N/A
Flow cytometry, violet laser
I could quantify PD1high CXCR5+ CD4 T follicular helper cells in the spleen and study their cytokine production
The antibody works well and gives bright signal, even for T cells
Can be difficult to compensate with other antibodies in the violet channel and some APC conjugated-antibodies
Very good for T follicular cell studies.