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Found 3 matching solutions for this experiment
| Upstream tips |
Protocol tips |
Downstream tips |
| For assessment of CD200 and CD200R1, we used anticoagulant in whole blood for a total volume of 200 μl per tube. After lysis by Red Blood Cell Lysis Buffer (Beijing Solarbio Science and Technology Co., Beijing, China), cells were resuspended in PBS. |
, tubes were incubated in the dark at room temperature for 30 min. After washing 2 times, fixing, and permeabilization, we added 10 μL CD200-PE (PE anti-mouseCD200, eBioscience, USA) and CD200R1-PE (PE anti-mouseCD200R1, eBioscience, USA), followed by mixing, and incubating for 30 min at room temperature. |
After washing, FACS Canto II flow cytometry (BD, USA) was performed. |
| Upstream tips |
| For assessment of CD200 and CD200R1, we used anticoagulant in whole blood for a total volume of 200 μl per tube. After lysis by Red Blood Cell Lysis Buffer (Beijing Solarbio Science and Technology Co., Beijing, China), cells were resuspended in PBS. |
| Protocol tips |
| , tubes were incubated in the dark at room temperature for 30 min. After washing 2 times, fixing, and permeabilization, we added 10 μL CD200-PE (PE anti-mouseCD200, eBioscience, USA) and CD200R1-PE (PE anti-mouseCD200R1, eBioscience, USA), followed by mixing, and incubating for 30 min at room temperature. |
| Downstream tips |
| After washing, FACS Canto II flow cytometry (BD, USA) was performed. |
| Upstream tips |
Protocol tips |
Downstream tips |
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Each cell type was single cell suspended and, incubated in FACS buffer containing pre-conjugated antibodies (see supplementary Table 2) for 20 min on ice. Following incubation, cells were washed twice with FACS buffer. |
All the flow cytometry performed using a Cell Lab Quanta SCMPL. The data were analyzed using Kaluza®1.2 software (Beckman Coulter, Denmark) and the expression of each CD markers on the cells was calculated based on the percentage and mean fluorescence intensity (MFI). |
| Protocol tips |
| Each cell type was single cell suspended and, incubated in FACS buffer containing pre-conjugated antibodies (see supplementary Table 2) for 20 min on ice. Following incubation, cells were washed twice with FACS buffer. |
| Downstream tips |
| All the flow cytometry performed using a Cell Lab Quanta SCMPL. The data were analyzed using Kaluza®1.2 software (Beckman Coulter, Denmark) and the expression of each CD markers on the cells was calculated based on the percentage and mean fluorescence intensity (MFI). |
| Upstream tips |
Protocol tips |
Downstream tips |
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Cells were incubated with antibodies in 0.1 M PBS (pH7.4) supplemented with 1% FCS and 0.1% sodium azide on ice for 30 minutes. Cells were then washed three times and fixed in 1% paraformaldehyde followed by flow cytometry analysis. |
Data were analyzed using the flowjo software (Tree Star, Inc., OR). |
| Protocol tips |
| Cells were incubated with antibodies in 0.1 M PBS (pH7.4) supplemented with 1% FCS and 0.1% sodium azide on ice for 30 minutes. Cells were then washed three times and fixed in 1% paraformaldehyde followed by flow cytometry analysis. |
| Downstream tips |
| Data were analyzed using the flowjo software (Tree Star, Inc., OR). |
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