One-step TUNEL In Situ Apoptosis Kit (Green, Elab Fluor® 488)
中文名称:一步法TUNEL原位细胞凋亡检测试剂盒(绿色,Elab Fluor® 488)
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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® 488 |
Ex/Em | 495/519 |
Channel set | FITC |
Filter set | FITC |
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-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:Is it avaliable to take TUNEL and IF assays together?
It is avaliable therometically, but it is recommanded to take a pre-test as there is no such data yet. You might need to notice some key points: 1. Protease K for section sample treatment as permeability agent may interfere the antigen of immunofluorescence; 2. High temperature or high pressure for antigen repair in immunofluorescence assay may lead to DNA fragmentation, that will end up with false positive TUNEL results.
Q2:Can the One-step TUNEL kit detect a sperm sample?
Yes, it can be used to test sperm samples.
Q3:How many samples can a 50 Assays kit afford?
A 50 assays kit can detect 50 samples with the size of about 5cm2 (the size of a cover slide or 12-well plate) without positive or negative control. It will take another two assays for control groups.
Q4:What is the difference between TUNEL in-situ kit and TUNEL flow cytometry kit?
The main difference lies in the detection methods, for TUNEL in-situ kits, a fluorescence microscope is used for observing the results, as for TUNEL flow ytometry kit, the sample will be detected by a flow cytometer. On the other hand, TUNEL in-situ kit is more suitable for tissue sections and cell slides or smears, while TUNEL flow ytometry kit is more suitable for suspended cells and adherent cells.
Q5:Is there any TUNEL assay kit that can detect apoptosis in erythrocyte?
The detection of TUNEL is based on DNA fragmentation in late stage apoptosis, so it is not avaliable for erythrocyte as there is no nuclei or other organelles containing DNA in erythrocyte.
Q6: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.
Q7:What is the well state of dewaxing?
The tissue is transparent, and no water drop can be left on the slide.
Q8:Can the cell sample be tested with the TUNEL kit without being prepared as smears and slides?
Yes, the cells can be collected into EP tubes as cell suspension. After being stained by following the procedure, the cells can be detected by flow cyrometer, or the suspension can be sucked onto a slide and observed with a microscope.
Q9: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.
Q10:For TUNEL assays, can a quenched slice be done again?
No. TUNEL works by attaching a fluorescently labeled dUTP to the 3'-OH terminal of the fragmented DNA, it can only be worked once since there is no 3'-OH terminal after the operation.