One-step TUNEL In Situ Apoptosis Kit (Red, Elab Fluor® 594)
中文名称：一步法TUNEL原位细胞凋亡检测试剂盒（红色，Elab Fluor® 594）
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- 应用： 细胞凋亡-DNA片段化
| 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 |
| 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®594 |
| Ex/Em | 590/617 |
| 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 |
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| E-CK-A211 | Annexin V-FITC/PI Apoptosis Kit | 200Assays , 100Assays , 50Assays , 20Assays |
| E-CK-A301 | Mitochondrial Membrane Potential Assay Kit (with JC-1) | 100Assays , 50Assays , 20Assays |
| E-CK-A321 | One-step TUNEL In Situ Apoptosis Kit (Green, Elab Fluor® 488) | 100Assays , 50Assays , 20Assays |
| E-CK-A362 | Enhanced Cell Counting Kit 8 (WST-8/CCK8) | 500Tests , 100Tests , 500Tests×20 |
Q1:Can frozen section be dried?
Do not dry frozen section. Frozen section can be stored in -20°C within a week or in -80°C for longer time after being prepared. Before the experiment, rewarm the section at room temperature for about 20 minutes, fix at room temperature for 30-60 minutes with 4% of paraformaldehyde formaldehyde, then wash it and follow the procedure on the manual.
Q2:Does TUNEL need antigen repair?
No, it doesn't. What's more, antigen repair may cause DNA fragmentation and lead to false positive signal.
Q3:Can I use neutral balsam for sealing?
DO NOT use neutral balsam. The section is in a water-soluble state after dyeing, while neutral balsam is a water-insoluble agent that will blurred the sample. It is recommended to use a water-soluble sealer.
Q4:What is the role of the TdT equilibration buffer in the TUNEL Kit?
The TdT equilibration buffer in the TUNEL kit is designed to provide optimal reaction conditions for the TdT enzyme.
Q5:What is the difference between TUNEL-fluorescence and HRP-DAB TUNEL?
Color development of DAB (result of TUNEL-HRP) is observed with an ordinary microscope, the results of TUNEL nuclear staining were served with white light. As for TUNEL fluorescence, the results of TUNEL and nuclear staining were observed with different fluorescence channels seperately with better definition and resolution.
Q6:Is it avaliable to take a TUNEL assay with a microplate instead of a cell slides/smears?
(1) The slides can be sealed with anti-fluorescence quenching agent, but the microplate cannot. (2) The amount of reagents will be increased with 6-well plate. (3) The visual field will be decreased due to the smaller diameter of wells.
Q7:Can TUNEL assay kit be used for mouse samples?
There is no species limination in TUNEL assay kits.
Q8:Is sealing agent containing anti-fluorescence quenching agent (including DAPI) able to be used in TUNEL?
Yes, it is. If DAPI is already contained in the anti-fluorescence quenching agent, there is no need to stain the sample with extra DAPI in the kit.
Q9:If the frozen section have been fixed before freezing, do they still need to be fixed and incubated at room temperature for 30min?
If it has been fixed, it is not necessary to incubate and fix again, and subsequent experimental steps can be directly carried out.
Q10:Is the cell slides/smears for TUNEL assay able to be stored overnight after fixation?
Yes, soak in PBS and store at 4℃.
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