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GFP-CERTIFIED® 细胞凋亡/坏死检测试剂盒
GFP-CERTIFIED® Apoptosis/Necrosis detection kit

GFP-CERTIFIED® 细胞凋亡/坏死检测试剂盒011593583977.png

GFP-CERTIFIED® Apoptosis/Necrosis detection kit



可进行区分正常细胞、早期凋亡细胞、晚期凋亡细胞和坏死细胞的多重检测,可与 GFP 和其他绿色荧光探针兼容。

 

● 可以兼容 GFP 和其他绿色荧光探针

● 容易区分正常细胞、早期凋亡细胞、晚期凋亡细胞和坏死细胞

● 已根据荧光显微镜和流式细胞仪等应用进行过优化

● 适用于死亡途径分析及药物、毒素研究

 

从凋亡到坏死的过渡是一个松散定义的连续体。质膜细胞外表面上磷脂酰丝氨酸(PS)的暴露仍是早期细胞凋亡的共同标志。这时候在Ca2+存在下,磷脂结合蛋白例如 Annexin V 与 PS 具有亲和力。由于 Annexin V 不具有细胞渗透性,所以这种胞外PS的结合是早期凋亡细胞对具有选择性的。

同样,质膜完整性的丧失,如不具有膜渗透性DNA嵌入染料若能标记出细胞核,则是表明晚期细胞凋亡和坏死的直接方法。考虑到Annexin V通常与荧光素(又称FITC)偶联,检测绿色荧光蛋白(GFP)表达细胞中的凋亡时可能会出现问题。以上两个荧光团的发射图谱会发生光谱重叠,使信号之间的区分变得困难或不可能,从而损害了总体上的数据质量。GFP-CERTIFIED® 细胞凋亡/坏死检测系统专为表达GFP的细胞系以及表达蓝色或青色荧光蛋白(BFP、CFPs)的细胞而设计,同时也可以应用于不带荧光蛋白的细胞的凋亡和坏死的细胞。另外,本试剂盒适合用于活细胞或结合了探针(如标记抗体、或与荧光素或香豆素显示相似光谱特性的其他荧光偶联物)的细胞。本试剂盒包含了用于确定细胞凋亡及坏死的早期和晚期阶段中所需的全部试剂。Annexin V -EnzoGold(增强型Cyanine-3)偶联物能够检测不同于荧光素或GFP的细胞凋亡。坏死检测试剂(红色)类似于发出红色的染料7-AAD,可促进晚期细胞凋亡和坏死检测。本试剂盒还包含了可用作阳性对照的凋亡诱导剂(Staurosporine)。检测试剂可单独使用或分别与其他单独或多重应用组合使用。本试剂盒还可以通过流式细胞仪和荧光/共焦显微镜提供快速、准确的检查结果。


ENZ-51002_Fig2.jpg


GFP、Annexin V-Cyanine 3 偶联物和坏死检测试剂(红色)的激发(阴影线)和发光(固体)光谱。

三种染料都可以轻易地被 488 nm 激光源激发。所有荧光团之间的发射峰值容易区分。


ENZ-51002_IF.jpg


GFP-CERTIFIED® 细胞凋亡/坏死检测试剂盒(ENZ-51002



ENZ-51002_Fig1A-B.jpg


流式细胞仪。用0.2%DMSO(panel A)模拟诱导 Jurkat 细胞,或用 2 μM 星形孢菌素(panel B)在37°C下诱导 Jurkat 细胞4h。经过处理后,在含有 Annexin V-Cyanine 3 和坏死检测试剂(红色;一种远红色发光的 DNA 嵌入染料)的缓冲液中孵育细胞,然后用 488 nm 激光的流式细胞仪进行分析,用 FL2(细胞凋亡)和 FL3(坏死)通道进行荧光检测。

模拟诱导的细胞凋亡和坏死主要呈阴性。处理4h 后,会出现3种细胞群:

(1)可以存活且不凋亡、不坏死的细胞(Annexin V-Cyanine 3、坏死检测试剂呈阴性);

(2)正在经历凋亡的细胞(Annexin V-Cyanine 3 呈阳性和 NDR 呈阴性);

(3)处于晚期凋亡和早期坏死的细胞(Annexin V-Cyanine 3 和坏死检测试剂呈阳性)。

 

ENZ-51002_Fig3.jpg

用 2.5 µM 星形孢菌素刺激 Jurkat 细胞 0-6 h。使用 Jurkat 悬浮细胞的流式细胞仪结果表现为早期凋亡(Annexin V-EnzoGold)和晚期凋亡/坏死(Annexin V-EnzoGold 和坏死染色)。



◆产品规格


 应用:

 流式细胞仪、荧光显微镜、荧光检测。

 使用事项/稳定性:

 Annexin V-EnzoGold 和结合缓冲液应在4°C储存。
 其他全部试剂应在-20°C储存。
 重组诱导剂(星形孢菌素)应在-20°C下储存。具体请参阅说明书。

 储存条件:

 避光。

 运输:

 蓝冰运输。

 短期储存温度:

 4°C

 试剂盒构成:

 凋亡检测试剂 (Annexin   V-EnzoGold)
 坏死检测试剂
 凋亡诱导剂(Staurosporine)
 结合缓冲液(10X)

 技术信息/产品说明:

 应用说明:
 Image-Based Analysis of a Human Neurosphere Stem Cell Model for the Evaluation of Potential 

 Neurotoxicants
 引用样本:
 For an overview on cited samples please click here.

 监管状况:

 RUO-仅供研究用



点击下载此处下载相关产品宣传页


参考文献


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High mobility group box 1 released from necrotic cells enhances regrowth and metastasis of cancer cells that have survived chemotherapy: Y. Luo, et al.; Eur. J. Cancer. 49, 741 (2013), Application(s): Fluorescence microscopy using CT26 cells, 摘要;


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59.

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产品列表
产品编号 产品名称 产品规格 产品等级 备注
ENZ-51002-25 GFP-CERTIFIED® Apoptosis/Necrosis detection kit
细胞凋亡/坏死检测试剂盒(荧光显微镜&流式)(GFP细胞系)
25 assays - -
ENZ-51002-100 GFP-CERTIFIED® Apoptosis/Necrosis detection kit
细胞凋亡/坏死检测试剂盒(荧光显微镜&流式)(GFP细胞系)
100 assays - -

相关产品
自噬抑制剂
产品编号 产品名称 产品规格 产品等级 备注
BML-AP502-0025 3-Methyladenine 25 mg - 3-甲基腺嘌呤,PI3激酶抑制剂
BML-CM110-0100 Bafilomycin A1 100 μg - 巴伐洛霉素,ATPase抑制剂
自噬激活剂
产品编号 产品名称 产品规格 产品等级 备注
BML-PE180-0001 Thapsigargin 1 mg - 毒胡萝卜素,ATPase抑制剂
BML-CC104-0010 Tunicamycin 10 mg - 衣霉素,GlcNAc磷酸转移酶抑制剂
ALX-550-084-G005 Carbamazepine 5 g - 卡马西平,镇痉剂和止痛剂
BML-SL100-0005 C2 Ceramide 5 mg - C2-神经酰胺,细胞渗透性神经酰胺类似
BML-A275-0005 Rapamycin 5 mg - 雷帕霉素,免疫抑制剂
BML-CA409-0050 Xestospongin C 50 μg - IP3受体抑制剂
自噬化合物库
产品编号 产品名称 产品规格 产品等级 备注
BML-AP502-0025 3-Methyladenine 25 mg - 3-甲基腺嘌呤,PI3激酶抑制剂
BML-CM110-0100 Bafilomycin A1 100 μg - 巴伐洛霉素,ATPase抑制剂

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