Calcein AM/PI Double Staining Kit

中文名称：Calcein AM/PI活死细胞双染试剂盒

货号：E-CK-A354
说明书

Price: ¥2000 ¥700 ¥300

Size:

2000Assays 500Assays 100Assays

应用： 细胞活力/细胞毒性

检测原理	Elabscience^® Calcein AM/PI Double Staining Kit可用于区分含有酯酶活性的哺乳动物的死细胞和活细胞，Calcein AM 是在传统的Calcein上添加上乙酰甲氧基甲酯即（AM）基团，疏水性增加，能很容易穿透活细胞膜，进入细胞内。Calcein AM本身无荧光，进入细胞后被细胞中内源性酯酶水解，生成具有强负电荷且不能通过细胞膜的极性分子Calcein滞留在细胞内，而Calcein可发出强绿色荧光（Ex/Em= 494nm/517nm）。由于死细胞缺乏酯酶，不能或很少能产生Calcein，因此仅活细胞会被染色为强绿色荧光，死细胞不能被染色或者染色非常弱。死细胞的细胞膜选择透过性丧失，碘化丙啶（Propidium Iodide，PI）可以进入胞内与双链DNA特异性结合并产生强烈的红色荧光（Ex/Em=535nm/617nm），从而对死细胞进行标记。因此，Calcein AM与碘化丙啶联合使用，可以对活细胞与死细胞同时进行双重荧光染色，可用于细胞活性与细胞毒性的检测。
检测方法	荧光;流式
样本类型	细胞
检测时间	30min
检测仪器	流式细胞仪;荧光显微镜
荧光	Calcein/PI
Ex/Em	494/517,535/617
Channel set	FITC, PE/PerCP/Cy5.5
Filter set	FITC, TRITC
自备试剂	PBS(E-BC-R187)
保存温度	该产品可在-20°C避光条件下保存12个月。
有效期	12个月
运输条件	冰袋

4T1 cells and Hela cells were treated with 5 % DMSO, 5μM Camptothecin, 10μM Camptothecin for 4h, then stained with Calcein-AM / PI Double Staining Kit and photographed.

Molt-4 cells were treated with 5 % DMSO and 10 % DMSO for 4h, then stained with Calcein-AM / PI Double Staining Kit and detected by flow cytometry.

Jurkat cells were placed at 4 °C for 20 days, then stained with Calcein-AM / PI Double Staining Kit and detected by flow cytometry.

E-CK-A153	Calcein AM Assay Buffer	100mL
E-CK-A164	Calcein AM Solution (100 μM)	500Tests , 100Tests , 500Tests×10
E-CK-A165	Propidium Iodide (PI) Solution(750 μM)	500Tests , 100Tests , 500Tests×10

Q1:What is the recommended storage condition (recommended minimum volume, buffer dissolution and concentration required and duration of solution storage) of calcein AM kit? Can the reagent be diluted to the used buffer concentration before being repackaged?

It is not recommended to dilute the dye before repackage, since it may lead to hydrolysis of Calcein AM as Calcein AM is more stable in stock solution. Calcein AM stock solution can be divided according to needs, and can be stored at -80°C for a longer shelf life. It is better to prepare the diluted buffer right before use, and use it up within one day.

Q2:Why it needn't to wash the samples of calcein staining before detection?

Calcein AM has strong fluorescence only after being hydrolyzed by endogenous esterase in the cell. Free calcein AM in buffer is almost non-fluorescent, so it does not need to be washed before detecting.

Q3:How many samples can you do with Calcein AM/PI Double Staining Kit from 100Assays?

Flow cytometry: 125 samples (1~5×10^5 cells) Fluorescence microscopy: 100 samples (96-well plate), 50 samples (24-well plate)