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上海善然生物科技有限公司
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Features and Benefits• Sensitive: The direct labeling procedure using fluorescein-dUTP reduces background labeling
• Fast: The use of fluorescein-dUTP allows analysis of the samples directly after the TUNEL reaction
• Convenient: No secondary detection system required
• Accurate: Identification of apoptosis at a molecular level (DNA-strand breaks) and identification of cells at the very early stages of apoptosis
General description下载特定产品批次和规格的CofA、SDS和IFU。 Kit for the detection and quantification of apoptosis at the single-cell level, based on labeling of DNA strand breaks (TUNEL technology); analysis by fluorescence microscopy or flow cytometry.Widely used methods to determine apoptosis include the analysis of the genomic DNA by agarose-gel electrophoresis and DNA fragmentation assays based on 3H-thymidine and, alternatively, 5-Bromo-2′-deoxy-uridine. The methods involve the separation of fragmented, low molecular weight DNA from unfragmented, high molecular weight DNA in a given cell population. Thus, these methods do not provide information about the fate of an individual cell in a given cell population, or particularly, in tissue sections. Alternatively, individual apoptotic cells may be microscopically recognized because of the characteristic appearance of nuclear chromatin condensation and fragmentation, but this method is subjective and limited to a relatively narrow time window when the morphological changes are at a maximum.
The hallmark of apoptosis is DNA degradation, which in early stages, is selective to the internucleosomal DNA linker regions. The DNA cleavage may yield double-stranded and single-stranded DNA breaks (nicks). Both types of breaks can be detected by labeling the free 3′-OH termini with modified nucleotides (e.g., biotin-dUTP, DIG-dUTP, fluorescein-dUTP) in an enzymatic reaction. The enzyme terminal deoxynucleotidyl transferase (TdT) catalyzes the template-independent polymerization of deoxyribonucleotides to the 3′-end of single- and double-stranded DNA. This method has also been termed TUNEL (TdT-mediated dUTP-X nick end labeling). Alternatively, free 3′-OH groups may be labeled using DNA polymerases by the template-dependent mechanism called nick translation. However, the TUNEL method is considered to be more sensitive and faster.Other Notes仅用于生命科学研究。不可用于诊断。包装1 kit containing 2 components.Preparation NoteWorking solution: Add total volume (50 μl) of Enzyme Solution to the remaining 450 μl Label Solution to obtain 500 μl TUNEL reaction mixture.
Mix well to equilibrate components.SpecificityThe TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. This allows discrimination of apoptosis from necrosis and from primary DNA strand breaks induced by cytostatic drugs or irradiation.ApplicationThe In Situ Cell Death Detection Kit, Fluorescein, is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level by fluorescence microscopy and quantitative detection by flow cytometry in cells and tissues.[5][6] Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems.
Examples are:
• Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research[3][4][7]
• Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research[1]
• Typing of cells undergoing cell death in heterogeneous populations by double staining procedures[2]
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文献和实验时样本之间干扰。 Q:说明书上写细胞量为 1~5 × 105 ,细胞量多的话对检测结果有没有影响? A:建议细胞量最多不要超过 106个,过量的细胞相当于染料被稀释,影响染色和分离效果。一般 2×105 个细胞足够进行凋亡检测了,通常情况下检测 10,000 个细胞就能得到比较好的结果。 Q:是否可以用 PBS 替代凋亡试剂盒里面的 Binding Buffer? A:不行,Binding Buffer 是必须要加的,Annexin V 与 PS 的结合依赖 Ca2+,Binding
【求助】Annexin V-FITC/PI细胞凋亡试剂盒的使用
mmsong1987 我买了Annexin V-FITC/PI细胞凋亡试剂盒,想研究某种材料的细胞毒性,看其是否会诱导细胞凋亡。之前做过MTT之类的检测,结果显示材料基本无毒性。 由于条件限制(流式细胞仪在我们学校另一个校区)不能及时进行检测,前两天进行了一次实验,是将细胞收集后拿到测试地点再进行的荧光染色处理,检测结果显示阴性对照组没有加材料处理的细胞也多处于凋亡状态,个人猜测可能跟细胞在路上耽误的时间有关(悬浮在Annexin V结合液中
【求助】急切盼知对Roche和Promega公司的TUNNEL凋亡试剂盒质量和信价比的评价
yfsm 最近想买TUNNEL凋亡试剂盒,代理商分别给我推荐了Roche和Promega公司,以前没做过心理没辙,想请各位专家帮忙分析一下,非常感谢! xiaoluks 我想做TNNEL,罗氏的给你报价多少钱啊,楼主想买哪个货号的,如果好的话我也买一样的,呵呵,我也是没做过的,不过当时师姐有用DAB显色法做的,是用的罗氏的试剂盒,质量挺好的,不过我想用荧光的,楼主有什么意见? woxingwosu
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