在成功收购德国宝灵曼公司（Boehringer Mannheim GmbH.）后，成立了专业服务于科学研究领域用户的罗氏诊断生命科学部（Roche Life Science），凭借半个多世纪在生命科学研究领域积累的经验，罗氏诊断生命科学部已成为在此领域中具重要地位的试剂及系统供应商之一。罗氏诊断生命科学部还为生命科学行业、生物制药行业和诊断行业提供所需的原料、试剂组分和试剂盒，罗氏诊断生命科学部为客户进行量身定制产品，满足客户个性化需求。
产品介绍
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
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 on3H-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 deoxynucleotidyltransferase (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-Xnickendlabeling). 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 Note
Working 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.
Specificity
The 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.
Application
TheIn SituCell 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.[4][5]Thus, theIn SituCell 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[7][3][]• Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research[10]• Typing of cells undergoing cell death in heterogeneous populations by double staining procedures[6]
