► 流式检测方案
【试剂】一、刺激剂和阻断剂
pma
multisciences,70-cs0001，溶于乙醇，0.1 mg/ml，100μl 包装。-20℃保存。
ionomycin
multisciences,70-cs0002，溶于乙醇，1 mg/ml，50μl 包装。-20℃保存。
monensin
multisciences,70-cs0004，溶于乙醇，50 mg/ml，100μl 包装。4℃保存至少4个月。
lps
二、固定破膜剂fix&perm kit（multisciences, 70-gas003）
三、fc 受体阻断剂人：human fc receptor binding inhibitor purified (ebioscience, 85-14-9161)
小鼠：anti-mouse cd16/cd32 (2.4g2), purified (multisciences, 70-am016)
四、细胞培养液rpmi 1640，含 10% fcs、200 μg/ml penicillin、200 u/ml streptomycin 和 4mml-glutamine
五、淋巴细胞分离液使用前恢复至室温。
六、b 细胞分选磁珠及检测试剂(可选)人：
cd19 microbeads, human (99-130-050-301)，检测抗体如 anti-human cd19 apc(ebioscience, 85-17-0199) 或 anti-human cd19 fitc (ebioscience, 85-11-0199) 或anti-human cd19 pe (ebioscience, 85-12-0199)等
小鼠：
cd19 microbeads, mouse (miltenyi, 99-130-052-201)，检测抗体如 anti-mousecd19 fitc (ebioscience, 85-11-0193) 或 anti-mouse cd19 pe (ebioscience, 85-12-0193)或anti-mouse cd19 apc (ebioscience, 85-17-0193)等；或 cd45r (b220) microbeads, mouse (miltenyi, 99-130-049-501)，检测抗体如anti-human/mousecd45r (b220) fitc (ebioscience,85-11-0452)或anti-human/mouse cd45r (b220) pe (ebioscience, 85-12-0452)或anti-human/mouse cd45r (b220) apc (ebioscience, 85-17-0452)等
七、抗体人：管号 抗体 厂商 目录号 注释
1 mouse igg2b κ isotype control pe ebioscience 85-12-4732 表面染色(对应于 cd1d)
1 mouse igg2a κ isotype control percp-cy5.5 ebioscience 85-45-4724 表面染色(对应于 cd5)
1 rat igg1 κ isotype control efluor® 660 ebioscience 85-50-4301 胞内染色(对应于 il-10)
2 anti-human cd19 fitc ebioscience 85-11-0199 表面染色
2 anti-human cd1d pe ebioscience 85-12-0016 表面染色
2 anti-human cd5 percp-cy5.5 ebioscience 85-45-0058 表面染色
2 anti-human il-10 efluor® 660 ebioscience 85-50-7108 胞内染色
小鼠：管号 抗体 厂商 目录号 注释
1 rat igg2b κ isotype control pe ebioscience 85-12-4031 表面染色(对应于 cd1d)
1 rat igg2a κ isotype control percp-cy5.5 ebioscience 85-45-4321 表面染色(对应于 cd5)
1 rat igg2b κ isotype control apc ebioscience 85-17-4031 胞内染色(对应于 il-10)
2 anti-mouse cd19 fitc ebioscience 85-11-0193 表面染色
2 anti-mouse cd1d pe ebioscience 85-12-0011 表面染色
2 anti-mouse cd5 percp-cy5.5 ebioscience 85-45-0051 表面染色
2 anti-mouse il-10 apc ebioscience 85-17-7101 胞内染色
八、流式染色缓冲液flo
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