TUNEL In Situ Apoptosis Kit (HRP-DAB Method)
中文名称：TUNEL原位细胞凋亡检测试剂盒（HRP-DAB显色法）
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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DNA内切酶，剪断核小体间的基因组DNA，暴露的3’-OH可在末端脱氧核酸转移酶（TdT）的催化下加上生物素标记的dUTP，辣根过氧化物酶(HRP)标记的Streptavidin (Streptavidin-HRP)可与之结合，在HRP的催化下通过DAB显色可观测到凋亡细胞，从而可通过普通光学显微镜检测到细胞的凋亡。由于正常的或正在增殖的细胞中几乎没有DNA的断裂，因而没有3'-OH形成，很少能够被标记。这就是TUNEL(TdT-mediated dUTP Nick-End Labeling)法检测细胞凋亡的原理。 |
| 检测方法 | DAB显色 |
| 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 | 普通光学显微镜 |
| 自备试剂 | 4%多聚甲醛(E-IR-R113),0.2% Triton X-100(E-IR-R122),3% H2O2(E-IR-R115),二甲苯，乙醇，DNase I，PBS(E-BC-R187),ddH2O,苏木素(E-IR-R120)，中性树胶(E-IR-R118) |
| 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-A320 | One-step TUNEL In Situ Apoptosis Kit (Green, FITC) | 100Assays , 50Assays , 20Assays |
| E-CK-A362 | Enhanced Cell Counting Kit 8 (WST-8/CCK8) | 500Tests , 100Tests , 500Tests×20 |
Q1:Can DNase I Bufferr be diluted with DEPC water?
It is recommended to dilute with ultra-pure water, DO NOT use DEPC water. DEPC water is used to protect RNA or DNA from enzymatic hydrolysis, it will deactive Dnase I. DEPC water can be used to dissolve RNA when extracting RNA.
Q2:What is the well state of dewaxing?
The tissue is transparent, and no water drop can be left on the slide.
Q3:Can TUNEL be used to observe apoptosis in Drosophila melanogaster tissues?
There is no species limination in TUNEL assay kits, so it will be avaliable theromatically. It is recommanded for customers to explore the method of sample preparation as there is no such data for insect samples.
Q4:What are the reasons that lead to false positives in TUNEL?
Cells with high proliferative or metabolic active may have a break in the DNA strand, both cases may end up with false-positive result. Evaluation of cell morphology may be used if there are doubts about the interpretation of the results.
Q5: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.
