One-step TUNEL In Situ Apoptosis Kit (Red, Elab Fluor® 555)

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中文名称：一步法TUNEL原位细胞凋亡检测试剂盒（红色，Elab Fluor® 555）

货号：E-CK-A325
文献（24）
说明书

说明书

One-step TUNEL In Situ Apoptosis Kit (Red, Elab Fluor® 555)

Price: ¥3600.00 ¥2000.00 ¥1500.00

Size:

100 Assays 50 Assays 20 Assays

应用： 细胞凋亡-DNA片段化

实验操作

检测原理	细胞在发生凋亡时，会激活一些特异性的DNA内切酶，这些内切酶会切断核小体间的基因组DNA。使得凋亡细胞的DNA被切割成180~200bp片段，在琼脂糖凝胶上通常以180~200bp的阶梯状迁移。TdT酶（脱氧核糖核酸末端转移酶）将标记的dUTP连接到断裂DNA暴露的3'-OH末端，通过在这些末端添加荧光dUTP的方式来标记晚期凋亡细胞，从而可以通过荧光显微镜进行检测。
检测方法	荧光法
样本类型	石蜡切片;冰冻切片;细胞爬片;细胞涂片
检测时间	3 hours
检测仪器	荧光显微镜
荧光	Elab Fluor®555
Ex/Em	555/565
Filter set	TRITC
自备试剂	4%多聚甲醛(E-IR-R113),0.2% Triton X-100(E-IR-R122),PBS(E-BC-R187),ddH2O,含抗荧光淬灭剂的封片液(E-IR-R119),二甲苯,无水乙醇
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

Paraffin embedded rat heart was treated with DNase I to fragment the DNA. DNA strand breaks showed intense fluorescent staining in DNase I treated sample (red). The cells were counterstained with DAPI (blue).This photo was taken by confocal microscope.

Paraffin embedded rat kidney was treated with DNase I to fragment the DNA. DNA strand breaks showed intense fluorescent staining in DNase I treated sample (red). The cells were counterstained with DAPI (blue).

Paraffin embedded mouse liver was treated with DNase I to fragment the DNA. DNA strand breaks showed intense fluorescent staining in DNase I treated sample (red). The cells were counterstained with DAPI (blue).

Fluorescence microscope analysis of camptothecin-induced apoptosis of Hela cells.

Mixed samples of normal mouse spleen cells and DNase I treated mouse spleen cells were stained.

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E-CK-A211	Annexin V-FITC/PI Apoptosis Kit	200 Assays , 100 Assays , 50 Assays , 20 Assays
E-CK-A301	Mitochondrial Membrane Potential Assay Kit (with JC-1)	100 Assays , 50 Assays , 20 Assays
E-CK-A321	One-step TUNEL In Situ Apoptosis Kit (Green, Elab Fluor® 488)	100 Assays , 50 Assays , 20 Assays
E-CK-A362	Enhanced Cell Counting Kit 8 (WST-8/CCK8)	500 Tests , 100 Tests , 500 Tests × 20

Q1:请问做tunel染色需要烤片吗？不烤片的话不会脱片吗？

制作石蜡切片的过程需要烤片，但是切片制作完毕之后做tunel染色的过程不需要烤片。

Q2:该如何给客户推荐合适的TUNEL原位凋亡试剂盒（荧光法）?

①实验样本不含荧光，所有的TUNEL试剂盒都可以使用。②如果实验样品带红光，推荐选用One-step TUNEL Assay Kit (Green, FITC) 货号：E-CK-A320或者One-step TUNEL Assay Kit (Green, Elab Fluor®488) 货号：E-CK-A320。
一般来说，Elab Fluor®488的抗淬灭效果优于FITC，如果需要长时间观察样本的绿色荧光推荐使用One-step TUNEL Assay Kit (Green, Elab Fluor®488)。
③如果实验样本带绿光，推荐选用One-step TUNEL Assay Kit (Red, Elab Fluor®594) 货号：E-CK-A322或者One-step TUNEL Assay Kit (Red, Elab Fluor®555) 货号：E-CK-A325。Elab Fluor®594与GFP的干扰更小，如果样本所带绿色荧光较强，优先推荐选择One-step TUNEL Assay Kit (Red, Elab Fluor®594) 货号：E-CK-A322。
④如果实验样品中同时有红色荧光和绿色荧光，推荐选用One-step TUNEL Assay Kit (Red, Elab Fluor®647) 货号：E-CK-A324，但是Elab Fluor®647对荧光显微镜要求比较高，需要显微镜配置有Cy5或Elab Fluor®647滤光片，一般共聚焦显微镜配置有该滤光片。

Q3:脱蜡干净的状态是什么样的呢？

组织透明，玻片不挂水即为脱蜡干净。

Q4:细胞样本没有做成涂片和爬片，而是直接固定渗透，可以用TUNEL试剂盒检测吗？

可以，客户可以将细胞收集到EP管中，按实验步骤进行后续操作，最后用流式细胞仪上机检测，或是吸取细胞悬液到载玻片上用显微镜观察。

Q5:组织切片封片可以使用中性树胶吗？

不能用中性树胶，因为切片染色完毕后，是处于水溶性状态，要用水溶性的封片剂，中性树胶是非水溶性封片剂，封片了之后会糊成一团，片子被破坏，用抗荧光淬灭封片剂来封片即可。

Q6:石蜡切片已经脱蜡通透了，能不能放到第二天再染色？

可以洗涤后浸泡在PBS中，放在4℃冰箱保存。

Q7:标记步骤中需要滴加多少体积的DAPI工作液呢？

DAPI染色时要求DAPI工作液覆盖样本即可。

Q8:有哪些原因会造成TUNEL假阳性呢？

具有高增殖或代谢活性细胞群，可能会存在DNA链的断裂，在这两种情况下，假阳性结果可能是样本本身带来的。当客户对结果的解释存疑时，细胞形态学的评价是一个重要的参数。