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Upstream tips |
Protocol tips |
Downstream tips |
A flow cytometer (LSR II; BD Biosciences) equipped with Diva V8.0 software was used for 9‐color phenotypic analysis. Laser alignment (405‐nm violet laser, 488‐nm blue laser, 640‐nm red laser) was verified with beads prior to running tumor cell samples |
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer. |
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Upstream tips |
A flow cytometer (LSR II; BD Biosciences) equipped with Diva V8.0 software was used for 9‐color phenotypic analysis. Laser alignment (405‐nm violet laser, 488‐nm blue laser, 640‐nm red laser) was verified with beads prior to running tumor cell samples |
Protocol tips |
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer. |
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