Intracellular Fixation/Permeabilization Buffer Kit
中文名称:胞内因子固定破膜剂

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保存温度 | 该产品可在 2~8°C条件下保存12个月。 |
有效期 | 12个月 |
运输条件 | 冰袋 |
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Q1:请问一下Th和Treg一个需要破胞膜一个破核膜,这两种细胞类型检测能一管做吗?
理论上可以一起做,但我们还是建议针对不同的实验选择相应的破膜剂。固定破核膜剂的破膜能力比较强,可以破核膜和胞膜,对核因子的检测效果更好,但可能会破坏胞膜上的一些特异性结合位点。固定破胞膜剂相对比较温和,适用于表达在细胞质上的抗原检测。
Q2:胞内固定破膜剂中破膜剂用细胞染色缓冲液稀释可以吗?
不行,建议使用超纯水、双蒸水、三蒸水或去离子水稀释。细胞染色缓冲液稀释破膜剂后会改变样本本身的渗透压以及盐粒子浓度,最终导致细胞状态变差甚至死亡。
Q3:组织样本检测胞内指标的注意事项有哪些?
①所有样本管均需与实验组一样进行固定破膜,如blank对照,单标组,fmo-iso;②各组固定破膜时间尽量不要相差太久,如果样本量多,建议分批次操作;③对于肿瘤组织样本,由于检测目的细胞量少,需考虑增加抗体用量,上机时间数至少100万个;④建议增加75um微米孔径滤网进行二次过滤,提高免疫细胞纯度和含量;⑤由于刺激阻断时间较长,需要在刺激阻断时间结束前注意无菌操作。
参考资料:
①血样胞内因子实验注意事项的直播回放:https://www.elabscience.cn/resource-flow_cytometry-detail-965。
②肿瘤组织单细胞悬液的制备方案:https://www.elabscience.cn/resource-flow_cytometry-detail-261。
Q4:流式抗体样本什么时候用无菌的耗材?
后续需要进行细胞培养操作的实验都需要使用无菌耗材,如胞内因子检测或流式分选实验。
Q5:我要染CD206,需要破膜,那我应该用细胞因子固定液还是转录因子固定液?
建议客户使用胞内因子固定破膜剂,货号为E-CK-A109。
