Calcein-AM是一种对活细胞进行荧光标记的细胞染色试剂,发绿色荧光(Ex=490 nm,Em=515 nm)。 因其在传统的Calcein(钙黄绿素)基础上引入乙酰甲氧基甲酯(AM)基团,增加了疏水性,使其能够轻易穿透活细胞膜。一旦进入细胞后,Calcein-AM(本身不发荧光)被细胞内的酯酶剪切形成膜非渗透性的极性分子Calcein,从而被滞留在细胞内并发出强绿色荧光。与其它同类试剂(如BCECF-AM和CFDA)相比,由于Calcein, AM细胞毒性极低,是最适合用于活细胞染色的荧光探针,而且不会抑制任何的细胞功能,如增殖和淋巴球的趋化性。
由于死细胞缺乏酯酶,Calcein, AM仅用于对活细胞的细胞生存能力测试和短期标记。作为核染色染料的碘化丙啶不能穿过活细胞的细胞膜,它穿过死细胞膜的无序区域而到达细胞核,并嵌入细胞的DNA双螺旋从而产生红色荧光(激发:535 nm,发射:617 nm),因此PI仅对死细胞染色。由于Calcein和PI-DNA都可被490 nm激发,因此可用荧光显微镜同时观察活细胞和死细胞。而用545 nm激发,仅可观察到死细胞。根据以上特点,Calcein, AM和PI经常被结合用来作为活细胞和死细胞的双重染色。由于不同细胞系的最佳染色条件不同,我们建议个别确定 Calcein, AM 和 PI的合适浓度。
本品为粉末形式提供的Calcein-AM,超纯级别(≥95%),可与碘化丙啶(PI)(Cat No. 40710ES03)联合使用,用于活细胞和死细胞的同时检测。或者直接购买太阳成集团提供的Calcein-AM/PI活细胞/死细胞双染试剂盒(Cat No. 40747ES76)。
中文名称(Chinese synonym) | 3'-O-乙酰胺-2',7'-二(羧乙基)-4或5-羧基荧光素,二乙酰甲酯;钙黄绿素乙酰甲酯; |
英文名称(English synonym) | 3',6'-Di(O-acetyl)-4',5'-bis[N,N-bis(carboxymethyl)aminomethyl]fluorescein, tetraacetoxymethyl ester |
CAS号( CAS NO) | 148504-34-1 |
分子式(Formula) | C46H46N2O23 |
分子量(Mol. Wt.) | 994.86 |
纯度(Purity) | ≥95%(HPLC) |
Ex/ Em(nm) | 490/515 |
结构式 | ![]() |
冰袋(wet ice)运输。-20℃干燥保存,避免直接暴露于强光下,有效期一年。
Q:这些探针染细胞,有没有细胞特异性?会不会 293 细胞不好染,MSC 好染?
A:由于这些染料是亲脂性染料,是通过插入到脂质膜中进而被激光激发观察,对于动植物细胞没有特异性。
Q:这些探针荧光探针装载完多久后观察?
A:为减少各种可能的误差,尽量缩短探针装载后到仪器测定所用的时间(刺激时间除外)。
Q:这些探针染色细胞后,对细胞有毒性吗?
A:荧光探针类产品对活细胞都会有一定的毒性,因此建议在染色处理完成后应尽快完成检测和观察。
Q:为什么染色后发现荧光颜色较浅,应怎么操作进行改进?
A:可以尝试增加染色的时间和浓度。
Q:要看几个小时过程中活细胞的形态变化,应该怎么选?
A:通常使用亲脂性花青染料,如 DiI,DiO,DiD 或 DiR。
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