Flow cytometrie analysisWe usually use Fisher tubes or equivalents as reaction tubesfor all steps described below.1)Wash the transfectant cells 3 times with washingbufferPBS containing 2% fetal calf serum (FCS)and 0.1%NaN;]. *Azide may react with copper or lead in plumbingsystem to form explosive metal azides. Therefore, always flushplenty of water when disposing materials containing azide intodrain.2)Resuspend the cells withwashingbuffer(4-6x105cells/mL).3)Add 400 uL of the cell suspension into each tube, andcentrifuge at 500 xg for 1 minute at room temperature(20~25°C). Remove supernatant by careful aspiration.4) Add 40 μL of the primary antibody at the concentration assuggested in the APPLICATIONS diluted in the washingbuffer. Mix well and incubate for 30-60 minutes at 4°C.
Elow细胞计数分析我们通常使用Fisher管或等效的反应管进行以下所有步骤。1）用含有2%小牛血清（FCS）和0.1%NaN的洗涤缓冲液（PBS）洗涤转染细胞3次。*叠氮化物可能与铜或铅在管道系统中反应形成爆炸性金属叠氮化物。因此，在处理含有叠氮化物的材料时，请务必用大量水冲洗排水系统。2）用洗涤缓冲液（4-6x105个细胞/mL）重新悬浮细胞。3）将400 μL细胞悬液加入每个管中，并在室温（20~25°C）下以500 xg离心1分钟。小心吸出上清液。4）加入40 μL的初级抗体，按照APPLICATIONS中建议的浓度稀释在洗涤缓冲液中。充分混合并孵育30-60分钟，温度为4°C。