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癌症生物学

Analysis of in vivo Interaction between RNA Binding Proteins and Their RNA Targets by UV Cross-linking and Immunoprecipitation (CLIP) Method 采用紫外线交联-免疫沉淀法(CLIP)分析RNA结合蛋白与其靶标RNA的体内互作 作者:Pamela Bielli and Claudio Sette日期:05/20/2017,浏览量:8200,Q&A: 0
RNA metabolism is tightly controlled across different tissues and developmental stages, and its dysregulation is one of the molecular hallmarks of cancer. Through direct binding to specific sequence element(s), RNA binding proteins (RBPs) play a pivotal role in co- and post-transcriptional RNA regulatory events. We have recently demonstrated that, ...
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Semi-quantitative Analysis of H4K20me1 Levels in Living Cells Using Mintbody 使用Mintbody半定量分析活细胞中H4K20me1的水平 作者:Yuko Sato and Hiroshi Kimura日期:05/20/2017,浏览量:7295,Q&A: 0
Eukaryotic nuclear DNA wraps around histone proteins to form a nucleosome, a basic unit of chromatin. Posttranslational modification of histones plays an important role in gene regulation and chromosome duplication. Some modifications are quite stable to be an epigenetic memory, and others exhibit rapid turnover or fluctuate during the cell cycle. ...
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Nucleosome Positioning Assay 核小体定位分析实验 作者:Zhongliang Zhao and Holger Bierhoff日期:05/20/2017,浏览量:9326,Q&A: 0
The basic unit of chromatin is the nucleosome, a histone octamer with 147 base pairs of DNA wrapped around it. Positions of nucleosomes relative to each other and to DNA elements have a strong impact on chromatin structure and gene activity and are tightly regulated at multiple levels, i.e., DNA sequence, transcription factor binding, ...
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Murine Bronchoalveolar Lavage 小鼠支气管肺泡灌洗 作者:Fan Sun, Gutian Xiao and Zhaoxia Qu日期:05/20/2017,浏览量:20687,Q&A: 1
A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting (FACS), PCR, bicinchoninic acid protein assay, enzyme-linked immunosorbent assay (ELISA), luminex assays, etc., to examine the immune cells, ...
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Isolation of Murine Alveolar Type II Epithelial Cells 小鼠肺泡II型上皮细胞分离 作者:Fan Sun, Gutian Xiao and Zhaoxia Qu日期:05/20/2017,浏览量:9092,Q&A: 0
We have optimized a protocol for isolation of alveolar type II epithelial cells from mouse lung. Lung cell suspensions are prepared by intratracheal instillation of dispase and agarose followed by mechanical disaggregation of the lungs. Alveolar type II epithelial cells are purified from these lung cell suspensions through magnetic-based negative ...
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Imaging the Pharynx to Measure the Uptake of Doxorubicin in Caenorhabditis elegans 咽部成像测定秀丽隐杆线虫中多柔比星的摄取 作者:Sivathevy Amirthagunabalasingam, Arturo Papaluca, Taramatti Harihar and Dindial Ramotar日期:05/20/2017,浏览量:6855,Q&A: 0
Caenorhabditis elegans offers an array of advantages to investigate the roles of uptake transporters. Herein, an epifluorescent microscopy approach was developed to monitor the uptake of the autofluorescent anticancer drug, doxorubicin, into the pharynx of C. elegans by organic cation transporters.
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细胞生物学

Flow Cytometric Analysis of Drug-induced HIV-1 Transcriptional Activity in A2 and A72 J-Lat Cell Lines A2和A72 J-Lat细胞系中药物诱导的HIV-1转录活性的流式细胞术分析 作者:Daniela Boehm and Melanie Ott日期:05/20/2017,浏览量:8999,Q&A: 0
The main obstacle to eradicating HIV-1 from patients is post-integration latency (Finzi et al., 1999). Antiretroviral treatments target only actively replicating virus, while latent infections that have low or no transcriptional activity remain untreated (Sedaghat et al., 2007). A combination of antiretroviral treatments with ...
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Phos-tag Immunoblot Analysis for Detecting IRF5 Phosphorylation Phos-tag免疫印迹分析检测IRF5磷酸化 作者:Go R. Sato, Tatsuma Ban and Tomohiko Tamura日期:05/20/2017,浏览量:11372,Q&A: 0
While the activation of the transcription factor interferon regulatory factor 5 (IRF5) is critical for the induction of innate immune responses, it also contributes to the pathogenesis of the autoimmune disease systemic lupus erythematosus (SLE). IRF5 phosphorylation is a hallmark of its activation in the Toll-like receptor (TLR) pathway, where ...
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免疫学

Lung Section Staining and Microscopy 肺切片染色和显微镜观察 作者:Xiaofeng Zhou and Bethany B Moore日期:05/20/2017,浏览量:18937,Q&A: 0
Our protocol describes immunofluorescent staining, hematoxylin and eosin staining and Masson’s trichrome staining on lung sections.
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Detection of ASC Oligomerization by Western Blotting 蛋白印迹法检测ASC寡聚化 作者:Jérôme Lugrin and Fabio Martinon 日期:05/20/2017,浏览量:14625,Q&A: 0
The apoptosis-associated speck-like protein with a caspase-recruitment domain (ASC) adaptor protein bridges inflammasome sensors and caspase-1. Upon inflammasome activation, ASC nucleates in a prion-like manner into a large and single platform responsible for the recruitment and the activation of caspase-1. Active caspase-1 will in turn promote ...
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Endogenous C-terminal Tagging by CRISPR/Cas9 in Trypanosoma cruzi 在克氏锥虫中利用CRISPR/Cas9内源标记靶基因C端 作者:Noelia Lander, Miguel A. Chiurillo, Aníbal E. Vercesi and Roberto Docampo日期:05/20/2017,浏览量:11737,Q&A: 0
To achieve the C-terminal tagging of endogenous proteins in T. cruzi we use the Cas9/pTREX-n vector (Lander et al., 2015) to insert a specific tag sequence (3xHA or 3xc-Myc) at the 3’ end of a specific gene of interest (GOI). Chimeric sgRNA targeting the 3’ end of the GOI is PCR-amplified and cloned into Cas9/pTREX-n vector. Then ...
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Creating a RAW264.7 CRISPR-Cas9 Genome Wide Library 创建RAW264.7 CRISPR-Cas9全基因组文库 作者:Brooke A Napier and Denise M Monack日期:05/20/2017,浏览量:10216,Q&A: 0
The bacterial clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 genome editing tools are used in mammalian cells to knock-out specific genes of interest to elucidate gene function. The CRISPR-Cas9 system requires that the mammalian cell expresses Cas9 endonuclease, guide RNA (gRNA) to lead the endonuclease to the gene of ...
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微生物学

Spore Preparation Protocol for Enrichment of Clostridia from Murine Intestine 用于小鼠肠道梭菌增菌培养的孢子制备方案 作者:Eric M. Velazquez, Fabian Rivera-Chávez and Andreas J. Bäumler日期:05/20/2017,浏览量:5969,Q&A: 0
In recent years, many spore-forming commensal Clostridia found in the gut have been discovered to promote host physiology, immune development, and protection against infections. We provide a detailed protocol for rapid enrichment of spore-forming bacteria from murine intestine. Briefly, contents from the intestinal cecum are collected aerobically, ...
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分子生物学

Assaying the Effects of Splice Site Variants by Exon Trapping in a Mammalian Cell Line 通过外显子捕获检测哺乳动物细胞系中剪接位点变体的作用 作者:Stuart W. Tompson and Terri L. Young日期:05/20/2017,浏览量:9574,Q&A: 2
There are several in silico programs that endeavor to predict the functional impact of an individual’s sequence variation at splice donor/acceptor sites, but experimental confirmation is problematic without a source of RNA from the individual that carries the variant. With the aid of an exon trapping vector, such as pSPL3, an investigator ...
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Ultradeep Pyrosequencing of Hepatitis C Virus to Define Evolutionary Phenotypes 超深度焦磷酸测序鉴定丙型肝炎病毒的进化表型 作者:Brendan A. Palmer, Zoya Dimitrova, Pavel Skums, Orla Crosbie, Elizabeth Kenny-Walsh and Liam J. Fanning日期:05/20/2017,浏览量:4816,Q&A: 0
Analysis of hypervariable regions (HVR) using pyrosequencing techniques is hampered by the ability of error correction algorithms to account for the heterogeneity of the variants present. Analysis of between-sample fluctuations to virome sub-populations, and detection of low frequency variants, are unreliable through the application of arbitrary ...
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CRISPR-PCS Protocol for Chromosome Splitting and Splitting Event Detection in Saccharomyces cerevisiae 检测酿酒酵母中染色体分裂和分裂事件的CRISPR-PCS实验方案 作者:Yu Sasano and Satoshi Harashima日期:05/20/2017,浏览量:8110,Q&A: 0
Chromosome engineering is an important technology with applications in basic biology and biotechnology. Chromosome splitting technology called PCS (PCR-mediated Chromosome Splitting) has already been developed as a fundamental chromosome engineering technology in the budding yeast. However, the splitting efficiency of PCS technology is not high ...
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A Method to Convert mRNA into a Guide RNA (gRNA) Library without Requiring Previous Bioinformatics Knowledge of the Organism 不需预知生物体生物信息学条件下将其mRNA转化为向导RNA(gRNA)库的方法 作者:Hiroshi Arakawa日期:05/20/2017,浏览量:7766,Q&A: 0
While the diversity of species represents a diversity of special biological abilities, many of the genes that encode those special abilities in a variety of species are untouched, leaving an untapped gold mine of genetic information; however, despite current advances in genome bioinformatics, annotation of that genetic information is incomplete in ...
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神经科学

Primary Olfactory Ensheathing Cell Culture from Human Olfactory Mucosa Specimen 人类嗅粘膜样本的原代嗅鞘细胞培养 作者:Mansoureh Hashemi and Mahmoudreza Hadjighassem日期:05/20/2017,浏览量:6280,Q&A: 0
The human olfactory mucosa is located in the middle and superior turbinates, and the septum of nasal cavity. Olfactory mucosa plays an important role in detection of odours and it is also the only nervous tissue that is exposed to the external environment. This property leads to easy access to the olfactory mucosa for achieving various researches. ...
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Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining H&E染色鉴定肌肉组织学特征并通过免疫荧光染色进行肌纤维分类 作者:Chao Wang, Feng Yue and Shihuan Kuang日期:05/20/2017,浏览量:20213,Q&A: 1
Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber ...
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A Tactile-visual Conditional Discrimination Task for Testing Spatial Working Memory in Rats 采用触觉-视觉条件辨别任务测试大鼠空间工作记忆 作者:Alicia Edsall, Zachary Gemzik and Amy Griffin日期:05/20/2017,浏览量:5361,Q&A: 0
This protocol describes a novel dual task comparison across two variants of a tactile-visual conditional discrimination (CD) T-maze task, one is dependent upon spatial working memory (SWM; CDWM) and the other one (CDSTANDARD) is not. The task variants are equivalent in their sensory and motor requirements and overt behavior of the rat. ...
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Locomotor Assay in Drosophila melanogaster 黑腹果蝇的运动分析 作者:Qingqing Liu, Jingsong Tian, Xing Yang, Yan Li and Aike Guo日期:05/20/2017,浏览量:7818,Q&A: 0
This protocol describes a simple locomotor assay in Drosophila melanogaster. In brief, the locomotor of each single fly in the culture dish is recorded by a web camera. The moving time, walking length, speed and the locomotor trails of the single fly could be quantitatively analyzed.
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Isolation and Cultivation of Primary Brain Endothelial Cells from Adult Mice 从成年小鼠分离和培养原代脑内皮细胞 作者:Julian Christopher Assmann, Kristin Müller, Jan Wenzel, Thomas Walther, Josefine Brands, Peter Thornton, Stuart M. Allan and Markus Schwaninger日期:05/20/2017,浏览量:10667,Q&A: 0
Brain endothelial cells are the major building block of the blood-brain barrier. To study the role of brain endothelial cells in vitro, the isolation of primary cells is of critical value. Here, we describe a protocol in which vessel fragments are isolated from adult mice. After density centrifugation and mild digestion of the fragments, ...
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植物科学

1-MCP (1-methylcyclopropene) Treatment Protocol for Fruit or Vegetables 水果或蔬菜的1-MCP(1-甲基环丙烯)处理实验方案 作者:Gamrasni Dan, Goldway Martin, Stern Yosi, Breitel Dario and Aharoni Asaph日期:05/20/2017,浏览量:13547,Q&A: 4
1-MCP (1-methylcyclopropene) is a simple synthetic hydrocarbon molecule that interacts with the ethylene receptor and inhibits the response of fruit or plant to ethylene. 1-MCP has opened new opportunities in handling harvested crops and serves as a powerful tool to learn about plant response to ethylene (Watkins and Miller, 2006). 1-MCP is ...
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Simple Spectroscopic Determination of Nitrate, Nitrite, and Ammonium in Arabidopsis thaliana 拟南芥中硝酸盐、亚硝酸盐和铵的简单光谱测定 作者:Takushi Hachiya and Yuki Okamoto日期:05/20/2017,浏览量:16059,Q&A: 2
Plants use nitrate, nitrite, and ammonium as inorganic nitrogen (N) sources. These N compounds are included in plant tissues at various concentrations depending on the balance between their uptake and assimilation. Thus, the contents of nitrate, nitrite, and ammonium are physiological indicators of plant N economy. Here, we describe a protocol for ...
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Exopolysaccharide Quantification for the Plant Pathogen Ralstonia solanacearum 植物病原体茄青枯病菌胞外多糖的定量分析 作者:Rémi Peyraud, Timothy P. Denny and Stéphane Genin日期:05/20/2017,浏览量:6545,Q&A: 0
Soluble exopolysaccharide is a major virulence factor produced by the plant pathogen Ralstonia solanacearum. Its massive production during plant infection is associated with the arrest of water flow in xylem vessels leading eventually to plant death. The composition of this heavy macromolecule includes mainly N-acetylgalactosamine. Here ...
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Plasma Membrane Preparation from Lilium davidii and Oryza sativa Mature and Germinated Pollen 兰州百合和亚洲水稻成熟和萌发花粉的浆膜制备 作者:Ning Yang, Bing Han, Lingtong Liu, Hao Yang and Tai Wang日期:05/20/2017,浏览量:6600,Q&A: 0
Pollen germination is an excellent process to study cell polarity establishment. During this process, the tip-growing pollen tube will start elongating. The plasma membrane as the selectively permeable barrier that separates the inner and outer cell environment plays crucial roles in this process. This protocol described an efficient aqueous ...
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Protein Isolation from Plasma Membrane, Digestion and Processing for Strong Cation Exchange Fractionation 用于强阳离子交换分馏的浆膜蛋白质的分离、消化和处理 作者:Ning Yang, Bing Han and Tai Wang日期:05/20/2017,浏览量:8640,Q&A: 0
Plasma membrane (PM) proteins play crucial roles in diverse biological processes. But their low abundance, alkalinity and hydrophobicity make their isolation a difficult task. This protocol describes an efficient method for PM proteins isolation, digestion and fractionation so that they can be well prepared for mass spectrometry analysis.
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干细胞

Efficient Production of Functional Human NKT Cells from Induced Pluripotent Stem Cells − Reprogramming of Human Vα24+iNKT Cells 诱导多能干细胞高效生成功能性人NKT细胞 - 人Vα24+iNKT细胞的重新编程 作者:Daisuke Yamada, Tomonori Iyoda, Kanako Shimizu, Yusuke Sato, Haruhiko Koseki and Shin-ichiro Fujii日期:05/20/2017,浏览量:9516,Q&A: 0
Antigen-specific T cell-derived induced pluripotent stem cells (iPSCs) have been shown to re-differentiate into functional T cells and thus provide a potential source of T cells that could be useful for cancer immunotherapy. Human Vα24+ invariant natural killer T (Vα24+iNKT) cells are subset of T cells that are characterized ...
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