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Found 3 matching solutions for this experiment
Upstream tips |
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Cells were stained with a fixable Live/Dead dye (ThermoFisher Scientific) concurrently with saturating concentrations of surface mAbs (Table S2) in a buffer containing 50%ABrilliant Violet Buffer (BD Bioscience) and 50%APBS for 30min at 4°C. |
Intracellular proteins were detected with saturating concentrations of mAbs for 30mins at 4°C in either a 0.1% saponin (SigmaAAldrich) buffer containing 10% FBS (SigmaAAldrich) or 1x PBS. All samples were acquired in a 0.1% saponin buffer on an X20FortessaASORP (BD Biosciences) and analysed in FlowJo (TreeStar). |
Protocol tips |
Cells were stained with a fixable Live/Dead dye (ThermoFisher Scientific) concurrently with saturating concentrations of surface mAbs (Table S2) in a buffer containing 50%ABrilliant Violet Buffer (BD Bioscience) and 50%APBS for 30min at 4°C. |
Downstream tips |
Intracellular proteins were detected with saturating concentrations of mAbs for 30mins at 4°C in either a 0.1% saponin (SigmaAAldrich) buffer containing 10% FBS (SigmaAAldrich) or 1x PBS. All samples were acquired in a 0.1% saponin buffer on an X20FortessaASORP (BD Biosciences) and analysed in FlowJo (TreeStar). |
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Human MSC (5 × 105 cells) were suspended in PBS and incubated with combinations of the monoclonal antibodies described above and also was acquired hMSC unstained used as control. |
A total of 100,000 events were acquired (at low speed). Data were analyzed using the Infinicyt program (Cytognos, Salamanca, Spain) |
Protocol tips |
Human MSC (5 × 105 cells) were suspended in PBS and incubated with combinations of the monoclonal antibodies described above and also was acquired hMSC unstained used as control. |
Downstream tips |
A total of 100,000 events were acquired (at low speed). Data were analyzed using the Infinicyt program (Cytognos, Salamanca, Spain) |
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Fluorescently labelled antibodies were validated for their sensitivity using monocyte derived macrophages |
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Protocol tips |
Fluorescently labelled antibodies were validated for their sensitivity using monocyte derived macrophages |
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