DetectionofBrdUIncorporationinDNASynthesizingCells
Detection of BrdU Incorporation in DNA Synthesizing Cells NOTE: Bromodeoxyuridine is a known carcinogen. Propidium iodine (PI) is known to be toxic and carcinogenic.1. Pulse actively growing cells in a tissue culture flask for one hour with 10 礛 BrdU (Sigma, Cat. No. B5002). 2. Pour contents of tissue culture flask into a centrifuge tube. Centrifuge 10 minutes at 400 x g (all cent......阅读全文
BrdU标记法检测原代细胞增殖
BrdU标记法检测原代细胞增殖1．细胞以1.5×105/ml细胞接于直径35ml培养皿中（内放置盖玻片），培养1天，用含0.4％ FCS培养液同步化3天，使绝数细胞处于G0期2．终止细胞培养，加入BrdU（终浓度30μg/L），37℃，孵育40min。3．弃培养液，玻片用PBS洗涤3次。4．甲醇/醋
Transfecting－Plasmid－DNA－into－NIH3T3－Cells－Using－Lipofectamine™－LTX－Reagent
实验概要Lipofectamine™ LTX Reagent is a proprietary, animal-origin free formulation for the transfection of DNA into eukaryotic cells with low cytotoxic
Isolation－and－Culture－of－human－vascular－smooth－muscle－cells
1. Human vascular smooth muscle cells (SMCs) were isolated from human saphenous vein.2. Small moistened pieces of tissue (6 mm2) were placed intima
其它PCR方法
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
原位PCR
About in situ PCR (Applied Biosystems)Basic information about in situ PCR and its applications.The In Situ PCR: Amplification and Detection in a Cellu
In－Situ－Cell－Death－(Apoptosis)－Detection－by－TUNEL－labeling
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. Beg
Staining－Procedure－for－Flow－Cytometric－Detection－of－Human－Cyclins
Staining Procedure for Flow Cytometric Detection of Human CyclinsThis is a standard protocol used at Pharmingen for Quality Control testing of the ant
Determination－and－Detection－of－Reactive－Oxygen－Species－(ROS),－Lipid－...
Reactive oxygen species or intermediates are formed by the incomplete reduction of oxygen. Organisms living in aerobic environment generate variou
Detection－Of－Cell－Viability－And/Or－Apoptosis－By－Flow－Cytometry－(FACS)
Viable cells are cells that when allowed to continue beyond the timepoint of examination will stay alive. Besides live and healthy cells, cells in ear
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement
IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance o
BrdU检测丁细胞和B细胞增殖
基本方案材 料实验动物0.8m g / m l B r d U (Sigma) 水 溶 液（口 服用）或 4m g / m l B r d U P B S 溶 液（注射用)P B S ， p H 7. 40.15mo l / L 冰冷的 NaCl冰 冷 的 9 5 % 乙醇多聚甲醛固定液D N A
BrdU标记法检测细胞增殖的原理
细胞增殖能力是测定物质毒性、评估药物安全、评价细胞活力的重要指标之一，被广泛应用于分子生物学、肿瘤生物学、免疫学和大规模药物筛选等研究领域。目前检测细胞增殖能力的方法主要分为两类：一类是直接测定进行分裂的细胞数来评价细胞增殖能力，如Ki67、BrdU以及EdU细胞增殖检测等；一类是通过测定活性细胞数
细胞周期测定实验——BrdU渗入法
实验方法原理细胞周期指细胞一个世代所经历的时间。从一次细胞分裂结束到下一次分裂结束为一个周期。细胞周期反应了细胞增殖速度。 单个细胞的周期测定可采用缩时摄影的方法，但它不能代表细胞群体的周期，故现多采用其他方法测群体周期。 BrdU（5-溴脱氧尿嘧啶核苷）加入培养基后，可做为细胞DNA复制的原料，经
Freezing－and－Thawing－cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
Transfecting－Suspension－Cells
实验概要将转移基因整合到细胞染色体DNA上,形成稳定表达转移基因的细胞系。 实验原理 细胞转染技术是目前广泛应用于病毒基因结构与功能以及基因调控等的研究。细胞转染可分为短暂转染和稳定(或永久) 转染两种。在短暂转染中,被转染基因并不整合至细胞染色体中,因而不能随细胞分裂而传代。转入病毒基因的转
Isolation－of－papillary－cells
实验概要This protocols provides a general protocol for isolation of papillary cells.实验步骤Isolation of renal papillary cells1. For isolation of papillary c
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read
Subculturing－Adherent－Cells
实验概要The following protocol describes a general procedure for subculturing adherent mammalian cells in culture.主要试剂1. Complete growth medium, pre-warme
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
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Isolation－of－papillary－cells
Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penicillin/
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB
细胞活性检测DNA合成增殖实验
BrdU检测法BrdU为胸腺嘧啶核苷（Thymine）的衍生物，可用于标记活细胞中新合成的DNA（细胞活性周期S期），BrdU可随着DNA复制进入子细胞，因此在加入BrdU后分裂增殖的细胞DNA都会带有BrdU标记，可通过抗BrdU单克隆抗体检测，借此检测细胞的增殖能力，但BrdU单克隆抗体检测过程
Caspase活化、核染色质固缩及DNA片段化
Life Technologies提供了用于细胞功能和健康检测的各种试剂。其中许多分析以荧光或比色法为基础，具有灵敏性、便利性和安全性。这些产品已经过多种仪器平台验证，包括显微镜、流式细胞仪、酶标仪和高内涵筛选，能够实现各种细胞功能的分析：细胞活性和细胞毒性细胞增殖和细胞周期细胞凋亡自噬氧化应激内吞
细胞增殖的检验方法：BrdU标记法
BrdU标记法1．细胞以1.5×105/ml细胞数接种于直径35ml培养皿中（内放置一盖玻片），培养1天，用含0.4％ FCS培养液同步化3天，使绝大多数细胞处于G0期。2．终止细胞培养前，加入BrdU（终浓度为30μg/L），37℃，孵育40min。3．弃培养液，玻片用PBS洗涤3次。4．甲醇/醋
细胞培养——细胞生长和细胞毒性
Articles posted in the Method Froum Cell Viability AssayDye exclusion method Viable Cell Counts Using Trypan Blue (Gibco) Soft Agar Assay For Colo
Isolation－of－rodent－pancreatic－β－cells
1. Adult rats weighing 250-350g were anesthetized, sacrificed and immediately used for pancreas sampling.2. Rat islets were isolated from male wistar
Decontamination－of－cells－from－the－yeast
I Destroy yeast1. Aspirate medium and wash cell in PBS.2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.3. In