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Found 3 matching solutions for this experiment
Upstream tips |
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Samples were isolated from mouse tissues or tumors as above or from trypsinized from tissue culture samples. |
Unstained samples and samples with antibody IgG controls were run in parallel. Additionally, color compensation for each fluorochrome was evaluated using IgG compensation beads (552843, BD Pharmingen) and ArC compensation beads (A10346, Invitrogen) with appropriate fluorescent marker controls. |
Flow cytometry was performed in the Stanford Flow Cytometry Core Facility using the BD Aria II (sorting) or BD LSR.II (analysis). |
Upstream tips |
Samples were isolated from mouse tissues or tumors as above or from trypsinized from tissue culture samples. |
Protocol tips |
Unstained samples and samples with antibody IgG controls were run in parallel. Additionally, color compensation for each fluorochrome was evaluated using IgG compensation beads (552843, BD Pharmingen) and ArC compensation beads (A10346, Invitrogen) with appropriate fluorescent marker controls. |
Downstream tips |
Flow cytometry was performed in the Stanford Flow Cytometry Core Facility using the BD Aria II (sorting) or BD LSR.II (analysis). |
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For c-kit+/SSEA1− cell isolation, cells were detached using HyQTase (Thermo Fisher Scientific), blocked with Fc (BioLegend, San Diego, CA, USA) for 15 min, and then incubated with the antibody at 4 °C for 30 min. Rinsing with staining buffer (eBioscience, San Diego, CA, USA). |
The cells were purified using a FACSAria Flow Cytometer (BD Bioscience). The purified cells were passaged five times before differentiation assays and cell transplantation. |
Protocol tips |
For c-kit+/SSEA1− cell isolation, cells were detached using HyQTase (Thermo Fisher Scientific), blocked with Fc (BioLegend, San Diego, CA, USA) for 15 min, and then incubated with the antibody at 4 °C for 30 min. Rinsing with staining buffer (eBioscience, San Diego, CA, USA). |
Downstream tips |
The cells were purified using a FACSAria Flow Cytometer (BD Bioscience). The purified cells were passaged five times before differentiation assays and cell transplantation. |
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Cultured cells were analyzed with FACS using anti-mouse CD19, CD3, and c-kit antibodies (BD Biosciences). |
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Cultured cells were analyzed with FACS using anti-mouse CD19, CD3, and c-kit antibodies (BD Biosciences). |
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