Miltenyi Biotec
Ki-67 Antibody, anti-human/mouse, REAfinity™
130-117-691
REA183
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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 FITC anti-ki67 antibody to detect proliferation in different cytokine-producing CD44+CD4 T cell subsets from old C57Bl/6J mice.
Flow Cytometry
Splenocytes of wild-type 12 months-old C57Bl/6J mouse, incubated in complete culture medium at 37 °C with PMA/ionomycin for 4h and brefeldin A for the last 3 hours.
Surface staining with anti-CD4 Al647 and CD44 BV785 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 to detect transcription factors (30 min at 4°C with fixative, and 15 min at 4°C with permeabilization buffer). Cells were incubated with FITC anti-Ki67 at 1/200 and different 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, blue laser
I could quantify Ki67 positive cells in relation with cytokine expression in the spleen.
The antibody works well, it is an excellent option to use in mouse compare to other anti-Ki67 antibodies rise in mouse and possibly interfering with anti-IgG staining to detect antibody producing cells.
None
Excellent option