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癌症生物学

Antisense Oligonucleotide-mediated Knockdown in Mammary Tumor Organoids 乳腺肿瘤类器官中反义寡核苷酸介导的基因敲低技术 作者:Sarah D. Diermeier and David L. Spector日期:08/20/2017,浏览量:7632,Q&A: 0
Primary mammary tumor organoids grown in 3D are an excellent system to study tumor biology. They resemble the organization and physiology of native epithelia more closely than cancer cell lines grown in 2D, and additionally model interactions with the ECM (Boj et al., 2015; Clevers, 2016; Shamir and Ewald, 2014). Mammary tumor organoids ...
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Mouse Model of Dextran Sodium Sulfate (DSS)-induced Colitis 葡聚糖硫酸钠(DSS)诱导的结肠炎小鼠模型 作者:Srustidhar Das, Surinder K. Batra and Satyanarayana Rachagani日期:08/20/2017,浏览量:14233,Q&A: 0
Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the intestinal tract and is primarily comprised of Crohn’s disease (CD) and ulcerative colitis (UC). Several murine models that include both chemical induced and genetic models have been developed that mimic some aspects of either CD or UC. These models have been instrumental in ...
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Rapid Profiling Cell Cycle by Flow Cytometry Using Concurrent Staining of DNA and Mitotic Markers DNA和有丝分类marker同步染色后利用流式细胞术快速分析细胞周期 作者:Yuqing Shen, Paolo Vignali and Ruoning Wang日期:08/20/2017,浏览量:9318,Q&A: 1
The flow cytometric quantitation of DNA content by DNA-binding fluorochrome, propidium iodide (PI) is the most widely used method for cell cycle analysis. However, the commonly used methods are time-consuming and labor-intensive and are incompatible with staining of mitotic markers by fluorescent-labeled antibodies. Here, we report an optimized ...
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Teratoma Formation Assay for Assessing Pluripotency and Tumorigenicity of Pluripotent Stem Cells 采用畸胎瘤形成分析法评估多能干细胞的多能性和致瘤性 作者:Shingo Miyawaki, Yohei Okada, Hideyuki Okano and Kyoko Miura日期:08/20/2017,浏览量:11990,Q&A: 0
Pluripotent stem cells such as induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) form teratomas when transplanted into immunodeficient mice. As teratomas contain all three germ layers (endoderm, mesoderm, ectoderm), teratoma formation assay is widely used as an index of pluripotency (Evans and Kaufman, 1981; Hentze et al. ...
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细胞生物学

Isolation and Analysis of Stromal Cell Populations from Mouse Lymph Nodes 小鼠淋巴结基质细胞群的分离和分析 作者:Yannick O. Alexandre and Scott N Mueller日期:08/20/2017,浏览量:8294,Q&A: 0
Our protocol describes a simple procedure for isolating stromal cells from lymph nodes (LN). LN are disrupted then enzymatically digested with collagenase and dispase to produce a single cell suspension that can be stained with fluorescently labelled antibodies and analysed by flow cytometry. This protocol will enable identification of ...
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Improved Oviduct Transfer Surgery for Genetically Modified Rat Production 用于转基因大鼠生产的改良输卵管移植手术 作者:Laura J. Lambert, Larry W. Johnson, Daniel Kennedy, Joan Cadillac and Robert A. Kesterson日期:08/20/2017,浏览量:7863,Q&A: 0
Rat embryo transfer surgeries are becoming more common with targeted nucleases increasing the demand for rat models. This protocol details pre-surgical preparation, improved surgical techniques for placing embryos into the oviduct, and post-surgical care of rats to parturition. Direct application of mouse oviduct transfer protocols results in ...
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免疫学

Isolation and Analysis of Stromal Vascular Cells from Visceral Adipose Tissue .内脏脂肪组织来源基质血管细胞的分离和分析 作者:Jessica Vu and Wei Ying日期:08/20/2017,浏览量:10453,Q&A: 0
The obesity epidemic is the underlying driver of the type 2 diabetes mellitus epidemic. A remarkable accumulation of various pro-inflammatory immune cells in adipose tissues is a hallmark of obesity and leads to pathogenesis of tissue inflammation and insulin resistance. Here, we describe a detailed protocol to isolate adipose tissue stromal ...
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Macrophage Survival Assay Using High Content Microscopy 使用高内涵显微技术进行巨噬细胞的生存分析 作者:Remzi Onur Eren and Nicolas Fasel日期:08/20/2017,浏览量:5765,Q&A: 0
Macrophages maintain tissue homoeostasis by regulating inflammation and tissue repair mechanisms. Thus, the fate of macrophages has an impact on the state of the tissue. The aim of this protocol is to quantify macrophage survival using high content microscopy and image processing software. Here, we describe a high-content image based protocol to ...
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Establishment of a Human Cell Line Persistently Infected with Sendai Virus 持续感染仙台病毒的人细胞系的建立 作者:Christopher Coakley, Cara Peter, Stephanie Fabry and Saurabh Chattopadhyay日期:08/20/2017,浏览量:6553,Q&A: 0
Interferon regulatory transcription factor 3 (IRF3) is a transcription factor that upon activation by virus infection promotes the synthesis of antiviral genes, such as the interferons (Hiscott, 2007). In addition to inducing genes, IRF3 triggers antiviral apoptosis by RIG-I-like receptor-induced IRF3 mediated pathway of apoptosis (RIPA), which is ...
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微生物学

Detection of Pathogens and Ampicillin-resistance Genes Using Multiplex Padlock Probes 使用多重锁式探针检测病原体和氨苄西林耐药基因 作者:Rick Conzemius and Ivan Barišić日期:08/20/2017,浏览量:6687,Q&A: 0
Diagnostic assays for pathogen identification and characterization are limited either by the number of simultaneously detectable targets, which rely on multiplexing methods, or by time constraints due to cultivation-based techniques. We recently presented a 100-plex method for human pathogen characterization to identify 75 bacterial and fungal ...
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Superoxide Dismutase (SOD) and Catalase (CAT) Activity Assay Protocols for Caenorhabditis elegans 秀丽隐杆线虫中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性测定实验方案 作者:Jing Zhang, Rui Chen, Zhenyang Yu and Lili Xue日期:08/20/2017,浏览量:31260,Q&A: 0
Assays for superoxide dismutase (SOD) and catalase (CAT) activities are widely employed to indicate antioxidant responses underlying the toxic effects of test chemicals. Yet, earlier studies mainly described the procedures as performed according to manufacturer’s instructions without modifications that are specific to any organisms. The present ...
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Snapshots of the Signaling Complex DesK:DesR in Different Functional States Using Rational Mutagenesis and X-ray Crystallography 利用合理化诱变和X射线晶体学分析在不同功能状态下的信号复合体DesK:DesR 作者:Juan Andres Imelio, Nicole Larrieux, Ariel Edgardo Mechaly, Felipe Trajtenberg and Alejandro Buschiazzo日期:08/20/2017,浏览量:5653,Q&A: 0
We have developed protocols to generate site-specific variants of the histidine-kinase DesK and its cognate response regulator DesR, conducive to trapping different signaling states of the proteins. Co-expression of both partners in E. coli, ensuring an excess of the regulator, was essential for soluble production of the DesK:DesR ...
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An Optimized Method for the Production Using PEI, Titration and Neutralization of SARS-CoV Spike Luciferase Pseudotypes 利用PEI优化生产,滴定和中和SARS-CoV spike萤光素酶假病毒的方法 作者:George Carnell, Keith Grehan, Francesca Ferrara, Eleonora Molesti and Nigel Temperton日期:08/20/2017,浏览量:12008,Q&A: 1
The protocol outlined represents a cost-effective, rapid and reliable method for the generation of high-titre viral pseudotype particles with the wild-type SARS-CoV spike protein on a lentiviral vector core using the widely available transfection reagent PEI. This protocol is optimized for transfection in 6-well plates; however it can be readily ...
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Protocol for HeLa Cells Infection with Escherichia coli Strains Producing Colibactin and Quantification of the Induced DNA-damage 用可产生大肠杆菌毒素的大肠埃希杆菌菌株感染HeLa细胞的实验方案及诱导产生DNA损伤的定量 作者:Nadège Bossuet-Greif, Marcy Belloy, Michèle Boury, Eric Oswald and Jean-Philippe Nougayrede日期:08/20/2017,浏览量:9662,Q&A: 0
Strains of Escherichia coli bearing the pks genomic island synthesize the genotoxin colibactin. Exposure of eukaryotic cells to E. coli producing colibactin induces DNA damages, ultimately leading to cell cycle arrest, senescence and death. Here we describe a simple method to demonstrate the genotoxicity of bacteria ...
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分子生物学

Assessment of Modulation of Protein Stability Using Pulse-chase Method 使用脉冲追踪法评估蛋白质稳定性的调节 作者:Mohamed Elgendy日期:08/20/2017,浏览量:9009,Q&A: 0
Pulse-chase technique is a method widely used to assess protein or mRNA stability. The principle of pulse-chase relies on labeling proteins or mRNA produced during a short period of time called ‘pulse’ and then following the rate of disappearance of those labeled proteins over a period of time called ‘chase’. This technique thus allows ...
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神经科学

A High-throughput Assay for mRNA Silencing in Primary Cortical Neurons in vitro with Oligonucleotide Therapeutics 体外使用寡核苷酸治疗药物对原代皮质神经元mRNA沉默的高通量测定 作者:Julia F. Alterman, Andrew H. Coles, Lauren M. Hall, Neil Aronin, Anastasia Khvorova and Marie-Cécile Didiot日期:08/20/2017,浏览量:8009,Q&A: 0
Primary neurons represent an ideal cellular system for the identification of therapeutic oligonucleotides for the treatment of neurodegenerative diseases. However, due to the sensitive nature of primary cells, the transfection of small interfering RNAs (siRNA) using classical methods is laborious and often shows low efficiency. Recent progress in ...
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Assessment of Thermal Pain Sensation in Rats and Mice Using the Hargreaves Test 使用Hargreaves测试法评估大鼠和小鼠的热疼痛感 作者:Menghon Cheah, James W Fawcett and Melissa R Andrews日期:08/20/2017,浏览量:10294,Q&A: 0
The Hargreaves test is specifically designed to assess thermal pain sensation in rodents such as rats and mice. This test has been used in experiments involving pain sensitization or recovery of thermal pain response following neural injury and regeneration. We present here a step-by-step protocol highlighted with important notes to guide ...
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An ex vivo Perifusion Method for Quantitative Determination of Neuropeptide Release from Mouse Hypothalamic Explants 用于定量测定小鼠下丘脑外植体神经肽释放的离体灌流方法 作者:Ophélia Le Thuc, Jacques Noël and Carole Rovère日期:08/20/2017,浏览量:6018,Q&A: 0
The hypothalamus is a primary brain area which, in mammals, regulates several physiological functions that are all related to maintaining general homeostasis, by linking the central nervous system (CNS) and the periphery. The hypothalamus itself can be considered an endocrine brain region of some sort as it hosts in its different nuclei several ...
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植物科学

Using Silicon Polymer Impression Technique and Scanning Electron Microscopy to Measure Stomatal Aperture, Morphology, and Density 使用硅聚合物印模技术和扫描电子显微镜技术测量气孔的开度、形态和密度 作者:Hui-Chen Wu, Ya-Chen Huang, Chia-Hung Liu and Tsung-Luo Jinn日期:08/20/2017,浏览量:7586,Q&A: 0
The number of stomata on leaves can be affected by intrinsic development programming and various environmental factors, in addition the control of stomatal apertures is extremely important for the plant stress response. In response to elevated temperatures, transpiration occurs through the stomatal apertures, allowing the leaf to cool through ...
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TUNEL Assay to Assess Extent of DNA Fragmentation and Programmed Cell Death in Root Cells under Various Stress Conditions 各种胁迫条件下根细胞DNA片段化和程序性细胞死亡的TUNEL法评估 作者:Amit K. Tripathi, Ashwani Pareek and Sneh Lata Singla-Pareek日期:08/20/2017,浏览量:11502,Q&A: 0
DNA damage is one of the common consequences of exposure to various stress conditions. Different methods have been developed to accurately assess DNA damage and fragmentation in cells and tissues exposed to different stress agents. However, owing to the presence of firm cellulosic cell wall and phenolics, plant cells and tissues are not easily ...
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ROS Detection in Botryococcus braunii Colonies with CellROX Green Reagent 使用CellROX绿色试剂检测布朗葡萄藻集落中的ROS 作者:Edmundo Lozoya-Gloria, Ivette Cornejo-Corona, Hem R. Thapa, Daniel R. Browne and Timothy P. Devarenne日期:08/20/2017,浏览量:6582,Q&A: 0
We analyzed the reactive oxygen species (ROS) accumulation in the colony-forming green microalga Botryococcus braunii in response to several stress inducers such as NaCl, NaHCO3, salicylic acid (SA), methyl jasmonate, and acetic acid. A staining assay using the fluorescent dye CellROX Green was used. CellROX Green is a ...
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Quantification of Membrane Damage/Cell Death Using Evan’s Blue Staining Technique 采用伊文思蓝染色技术定量膜损伤/细胞死亡 作者:Preethi Vijayaraghavareddy, Vanitha Adhinarayanreddy, Ramu S Vemanna, Sheshshayee Sreeman and Udayakumar Makarla 日期:08/20/2017,浏览量:15605,Q&A: 0
Membrane damage is a hallmark of both biotic and abiotic stress responses. The membrane determines the ability of a cell to sustain altered environmental conditions and hence can be used as a biomarker to assess stress-induced cell damage or death. We present an easy, quick, cost-effective, staining and spectrophotometric method to assess membrane ...
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干细胞

Exit from Pluripotency Assay of Mouse Embryonic Stem Cells 小鼠胚胎干细胞退出多能性测定实验 作者:Daniel Cirera-Salinas and Constance Ciaudo日期:08/20/2017,浏览量:6845,Q&A: 0
A novel method to assess the dissolution of the core pluripotency transcription-factor circuit of mouse Embryonic Stem Cells (mESCs) has been developed (Ying et al., 2003; Betschinger et al., 2013). In order to efficiently identify genes essential for the break-down of the pluripotency network in mutant mESCs with proliferation ...
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A Co-culture Assay to Determine Efficacy of TNF-α Suppression by Biomechanically Induced Human Bone Marrow Mesenchymal Stem Cells 人骨髓间充质干细胞共培养实验测定生物力学诱导的TNF-α抑制功效 作者:Miguel F. Diaz, Siobahn M. Evans, Scott D. Olson, Charles S. Cox Jr and Pamela L. Wenzel日期:08/20/2017,浏览量:6493,Q&A: 0
The beneficial effects of mesenchymal stem cell (MSC)-based cellular therapies are believed to be mediated primarily by the ability of MSCs to suppress inflammation associated with chronic or acute injury, infection, autoimmunity, and graft-versus-host disease. To specifically address the effects of frictional force caused by blood flow, or wall ...
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Isolation and Expansion of Mesenchymal Stem Cells from Murine Adipose Tissue 从鼠脂肪组织中分离和扩增间充质干细胞 作者:Natalia V. Andreeva, Alexandra A. Dalina and Alexander V. Belyavsky日期:08/20/2017,浏览量:9845,Q&A: 1
Mesenchymal stem cells (MSCs) are currently intensively studied due to significant promise which they represent for successful implementations of future cell therapy clinical protocols. This in turn emphasizes importance of careful preclinical studies of MSC effects in various murine disease models. The appropriate cell preparations with ...
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