InSituCellDeath(Apoptosis)DetectionbyTUNELlabeling
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Begin chilling Triton/SSC on ice.0.1% Triton/ 0.1% Sodium Citrate, 2 min., 4°C.All slides: 1x PBS rinse, 2 times (+ 10 min for those non-pos.control slides).(Pos. control slide: in DNase I solution (100µl of 200µg/ml), 10 min., RT. 1x PBS rinse, 2 times in a separate conta......阅读全文
同济大学毛志勇教授发表Cell-Death--Differentiation文章
基因组稳定性下降是生物体衰老发生极其重要的一个标志。细胞长期在各种因素的影响下,DNA遭受着多种损伤,若这些损伤不被及时准确地修复将诱发基因组稳定性的下降,进而影响细胞的正常生命活动。这些损伤中,DNA双链断裂(DSBs)是最为严重的基因组损伤之一。近年来,虽然关于DNA DSBs修复与衰老发
Cell-Death果蝇DNA损伤后细胞增殖与凋亡的协调
暴露于遗传毒性应激可促进细胞周期停滞和DNA修复或凋亡。这些“生”或“死”细胞命运的决定通常取决于肿瘤抑制基因53的活性。因此,对p53的精确调控对于维持组织内环境平衡和防止肿瘤的发展至关重要。然而,细胞周期进程是如何影响p53细胞命运的,目前尚不清楚。 已经提出了几种机制来解释dna损伤细胞
发育生物学
In Vitro Production of Bovine Embryos (P.J. Hansen Lab, Dept. of Animal Sciences, University of Florida) This protocol describe procedures for in v
细胞凋亡检测方法简介
细胞凋亡是指细胞在生长到一定阶段及某些生理过程中,会受到多因素和机制的严格调控而进入死亡。这是一个动态的过程,会涉及一系列复杂的生化反应, 是一个需要酶参与的级联反应,同时也涉及不同基因的表达及调控、信号传导。其中有几项生理过程能被用来检测凋亡的发生,因此多参数分析法对于准确检测这一过程十分
TUNEL-方法
Terminal deoxynucleotidyl Transferase-mediated dUTP nick end labeling (TUNEL) is an in situ method for detecting the 3'-OH ends of DNA exposed dur
BrdU-Labeling-Protocol
实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
Preparation-of-nucleic-acid-probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
Coenzyme-A-Detection
实验概要The experiment provided an easy, convenient assay to measure the CoA level in a variety of biological samples. In the assay, free CoA is specif
细胞周期的流式细胞伩检测实验方法(PI,Brdu)1
ANALYSIS OF CELL CYCLE Miriam Capri and Daniela Barbieri Dept. Biomedical Sciences, Sect. General Pathology,Via Campi, 287, University of Modena, 4110
In-situ-hybridization
Note: Although it is possible to perform in situ hybridization on bleached embryos, it appears to reduced the strength of the signal. For best results
CMFDA-Labeling-of-Platelet
OUTLINE CMFDA (5-chloromethylfluorescein diacetate) is a lipophilic tracer that has an enormous advantage over ordinary tracers (e.g. FITC) because
Arachidonic-Acid-Labeling
1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum. 2) Pellet cells and wash 1 time with room temperature PBS. 3) Resus
Immunofluorescence-Labeling-of-Cells
实验概要Antibodies are an important tool for demonstrating both the presence and the subcellular localization of an antigen. Cell staining is a very ver
Cell-Death-Differ:启动肺癌细胞自毁开关的新疗法
近日,英国癌症研究中心科学家们发现了一种药物组合,可以触发肺癌细胞的自毁过程,此项研究为新型治疗方法开发铺平了道路。 当健康细胞对机体不再有用,它们便引发一连串事件最终导致自我毁灭。但癌细胞则突然转向远离这个“自杀道路”,成为不朽,这意味着细胞生长失控,引起肿瘤的形成。 现在,英国癌症研究中
Cell-Death--Disease免疫、干细胞与疾病-国际研讨会召开
10月12日至14日,第五届Cell Death & Disease免疫、干细胞与疾病国际研讨会在中国常州市明都真儒酒店会议厅召开。常州市副市长张云云、卫生局党委书记、局长王莉、常州市第一人民医院院长何小舟、中国科学院上海生命科学研究院健康科学研究所主持工作的副所长孔祥银等领导出席会议开幕式并讲
第一届Cell-Death-and-Disease研讨会会议通知
第一届Cell Death and Disease研讨会暨第四届中英“细胞死亡、干细胞与癌症”国际研讨会将于2013年5月8日~9日在上海召开。 本次会议主题为:细胞死亡、干细胞与免疫,将邀请海内外相关领域卓有建树的科学家,采用大会报告及专题讨论的形式,结合国际上癌症生物学研究领域的前
RNA-FISH-on-cultured-cells-in-interphase
IntroductionFluorescence in situ hybridization (FISH) has become a widely used method in genome and molecular genetic studies. The technique is highly
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
A-rapid,-quantitative-and-inexpensive-method-for-detecting-apoptosis2
Figure 1.Determination of apoptosis in transiently transfected murine [beta] tumor cells. (A) The number of apoptotic [beta]HC 13T tumor cells (% apop
cell-death--disease:-黑色素瘤抗坏死效应分子机制
RIPK1与RIPK3被发现是参与细胞坏死性凋亡的关键成员,在caspase家族活性被抑制的情况下,RIPK1激活后能够与下游的RIPK3结合形成坏死小体,帮助RIPK3激活(磷酸化)。磷酸化后的RIPK3再次将MLKL磷酸化,磷酸化后的MLKL能够在细胞膜内侧自聚合形成寡聚体,最终导致细胞坏死
Cell-Death-and-Disease:-RNF128有望成为ALI药物干预的候选药物
急性肺损伤(ALI)和急性呼吸窘迫综合征(ARDS)是以严重的肺部炎症、肺泡毛细血管屏障受损和肺水肿为特征的危及生命的呼吸系统疾病,死亡率高。ALI发生发展的分子机制尚未完全阐明,但肺泡巨噬细胞和中性粒细胞在ALI的发病机制中起着关键作用。 激活的中性粒细胞和肺泡巨噬细胞释放过量的促炎细胞因子
Detection-and-Measurement-of-Radioactivity
Radioactive Decay Isotopes of a given element have nuclei with the same number of protons but different numbers of neutrons. Some isotopes are st
Detection-of-Glycoproteins-on-Blot
Detection of Glycoproteins on BlotSource: Contributed by Sharad Purohit, Paller's LabReagentsSodium acetate Buffer (200mM, pH 5.5): Prepare a 200
Detection-of-Mycoplasma-by-Culture
AimDetection of mycoplasma by culture is the reference method of detection and has a theoretical level of detection of 1 colony-forming unit (cfu). Ho
Detection-and-Measurement-of-Radioactivity
Liquid scintillation countingThe amount of kinetic energy in a beta particle differs from one decay to the next. However, each radioisotope has a typi
线粒体荧光探针大全:TMRM,Mitotracker,JC1(5)
Monoclonal Antibodies Specific for Proteins in the Oxidative Phosphorylation SystemOxidative phosphorylation (OxPhos) activity occurs in the mitochond
六种染色后光镜观察法检测肝癌细胞凋亡
作者:杨连君,司晓辉,王文亮,王文勇,赵一岭,方正清[摘 要] 目的:探讨简便易行的在光镜下通过形态学观察检测细胞凋亡的方法,并对其进行比较。方法:首先用6%的乙醇作用6 h诱导人肝细胞癌细胞系HCC9204细胞凋亡,然后进行未固定细胞的台盼蓝染色、吖啶橙/溴化乙啶(AO/EB)双染色,细胞固定后
DNA-labeling-by-nick-translation
DNA labeling by nick translationreagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, co
Cell-death-&diffe:CR1调控结肠癌肿瘤干细胞功能
近日,国际期刊cell death&differentiation 在线发表了意大利科学家的一项最新研究进展,他们发现胚胎期蛋白Cripto-1(CR1)在结肠癌肿瘤干细胞中表达能够促进肿瘤干细胞的克隆形成,这项研究表明CR1可能通过促进结肠癌肿瘤干细胞生长导致结肠癌复发。 最近,细胞干
流式细胞术定量检测细胞凋亡的3种方法研究比较
细胞凋亡的定性检测依赖于形态学观察及DNA电泳,其定量检测则需借助于流式细胞检查。使用碘化丙啶(PI)染色检测DNA含量是最早出现的凋亡定量检测方法[1]。进入90年代,检测DNA断裂点的TUNEL(Terminal deoxylnucleotidyl transferase mediated-dU