CD200 Monoclonal Antibody (OX90), PE, eBioscience™

Protocol tips

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.

Publication protocol

We collected blood from the orbits of the mice before treatment, 7 days after treatment, and 15 days after treatment. 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. We added 10 μL CD38-FITC (FITC anti-mouse CD38, Bioscience, USA) and 10 μL CD15-FITC (FITC anti-mouse). After mixing, 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.

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