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癌症生物学

Isolation and Separation of Epithelial CD34+ Cancer Stem Cells from Tgfbr2-deficient Squamous Cell Carcinoma 从Tgfbr2缺乏鳞状细胞癌分离上皮CD34+癌干细胞 作者:Heather A. McCauley and Géraldine Guasch日期:09/05/2017,浏览量:6946,Q&A: 0
Most epithelial tumors have been shown to contain cancer stem cells that are potentially the driving force in tumor progression and metastasis (Kreso and Dick, 2014; Nassar and Blanpain, 2016). To study these cells in depth, cell isolation strategies relying on cell surface markers or fluorescent reporters are essential, and the isolation ...
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GFP-Grb2 Translocation Assay Using High-content Imaging to Screen for Modulators of EGFR-signaling 通过高含量成像观测GFP-Grb2易位筛查EGFR信号转导调控因子 作者:Julia Petschnigg and Robin Ketteler日期:09/05/2017,浏览量:6046,Q&A: 0
High-content screening is a useful tool to understand complex cellular processes and to identify genes, proteins or small molecule compounds that modulate such pathways. High-content assays monitor the function of a protein or pathway by visualizing a change in an image-based readout, such as a change in the localization of a reporter protein. ...
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Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures 在细胞培养中建立基于多重图像的自噬RNAi筛选方法 作者:Jennifer Jung and Christian Behrends日期:09/05/2017,浏览量:6610,Q&A: 0
Autophagy is a recycling pathway, in which intracellular cargoes including protein aggregates and bacteria are engulfed by autophagosomes and subsequently degraded after fusion with lysosomes. Dysregulation of this process is involved in several human diseases such as cancer or neurodegeneration. Hence, advancing our understanding of how autophagy ...
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细胞生物学

Peroxisome Motility Measurement and Quantification Assay 过氧化物酶体能动性测量和定量分析 作者:Jeremy Metz, Inês G. Castro and Michael Schrader日期:09/05/2017,浏览量:5739,Q&A: 0
Organelle movement, distribution and interaction contribute to the organisation of the eukaryotic cell. Peroxisomes are multifunctional organelles which contribute to cellular lipid metabolism and ROS homeostasis. They distribute uniformly in mammalian cells and move along microtubules via kinesin and dynein motors. Their metabolic cooperation ...
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Staining of Membrane Receptors with Fluorescently-labeled DNA Aptamers for Super-resolution Imaging 用荧光标记的DNA吸附剂染色膜受体以实现超分辨率成像 作者:Maria Angela Gomes de Castro, Claudia Höbartner and Felipe Opazo日期:09/05/2017,浏览量:6888,Q&A: 3
One of the most prominent applications of fluorescent super-resolution microscopy is the study of nanodomain arrangements of receptors and the endocytic pathway. Staining methods are becoming crucial for answering questions on the nanoscale, therefore, the use of small and monovalent affinity probes is of great interest in super-resolution ...
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发育生物学

Labelling HaloTag Fusion Proteins with HaloTag Ligand in Living Cells 在活细胞中使用HaloTag配体标记HaloTag融合蛋白 作者:Huy Nguyen Duc and Xiaojun Ren 日期:09/05/2017,浏览量:8698,Q&A: 0
HaloTag has been widely used to label proteins in vitro and in vivo (Los et al., 2008). In this protocol, we describe labelling HaloTag-Cbx fusion proteins by HaloTag ligands for live-cell single-molecule imaging (Zhen et al., 2016).
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Isolation of Mouse Cardiac Neural Crest Cells and Their Differentiation into Smooth Muscle Cells 小鼠心脏神经嵴细胞的分离及向平滑肌细胞的分化 作者:Xia Wang and Sophie Astrof日期:09/05/2017,浏览量:6817,Q&A: 0
Cardiac neural crest cells (CNCCs) originate at the dorsal edge of the neural tube between the otic pit and the caudal edge of the 3rd somite, and migrate into the pharyngeal arches and the heart. We have shown that fibronectin (Fn1) plays an important role in the development of the CNCC by regulating the differentiation of CNCCs into ...
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免疫学

In vivo Priming of T Cells with in vitro Pulsed Dendritic Cells: Popliteal Lymph Node Assay 使用体外脉冲树突状细胞体外致敏T细胞: 腘窝淋巴结实验 作者:Songjie Cai, Masayuki Fujino, Lina Lu and Xiao-Kang Li日期:09/05/2017,浏览量:8946,Q&A: 0
One-way mixed lymphocyte reaction (MLR) is a classic tool to measure how T cells react to external stimuli. However, MLR is an in vitro reaction system, which shows different response intensity compared with in vivo trails sometimes due to the lack of cytokines, tissue matrix and other immune response associated factors. The ...
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微生物学

Sterol Analysis in Kluyveromyces lactis 乳酸克鲁维酵母中的甾醇分析 作者:Yvetta Gbelska, Nora Toth Hervay, Marcela Morvova and Alexandra Konecna日期:09/05/2017,浏览量:5429,Q&A: 0
Sterols are essential lipids of most eukaryotic cells with multiple functions (structural, regulatory and developmental). Sterol profile of yeast cells is often determined during the studies of ergosterol synthesis mutants used to uncover a number of functions for various sterols in yeast cells. Molecular studies of ergosterol biosynthesis have ...
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Determination of Local pH Differences within Living Salmonella Cells by High-resolution pH Imaging Based on pH-sensitive GFP Derivative, pHluorin(M153R) 采用基于pH敏感性GFP衍生物pHluorin(M153R)的高分辨率pH成像测定活体沙门氏菌细胞中局部pH值的差异 作者:Yusuke V. Morimoto, Nobunori Kami-ike, Keiichi Namba and Tohru Minamino日期:09/05/2017,浏览量:5443,Q&A: 0
The bacterial flagellar type III protein export apparatus is composed of a transmembrane export gate complex and a cytoplasmic ATPase complex. The export apparatus requires ATP hydrolysis and the proton motive force across the cytoplasmic membrane to unfold and transport flagellar component proteins for the construction of the bacterial flagellum ...
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Rearing of Culex spp. and Aedes spp. Mosquitoes 库蚊和伊蚊的饲养 作者:Elizabeth Kauffman, Anne Payne, Mary A. Franke, Michael A. Schmid, Eva Harris and Laura D. Kramer日期:09/05/2017,浏览量:11682,Q&A: 0
Mosquito-transmitted pathogens cause major public health problems and contribute substantially to the global burden of disease. Aedes mosquitoes transmit dengue, Zika, yellow fever, and Chikungunya viruses; Culex mosquitoes transmit West Nile, Japanese encephalitis, and Saint Louis encephalitis viruses, among others. Experiments ...
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Multicolor Stimulated Emission Depletion (STED) Microscopy to Generate High-resolution Images of Respiratory Syncytial Virus Particles and Infected Cells 利用多色受激发射损耗(STED)显微镜获取呼​​吸道合胞体病毒颗粒和感染细胞的高分辨率图像 作者:Masfique Mehedi, Margery Smelkinson, Juraj Kabat, Sundar Ganesan, Peter L. Collins and Ursula J. Buchholz日期:09/05/2017,浏览量:7040,Q&A: 0
Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these ...
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Expression and Purification of a Mammalian P2X7 Receptor from Sf9 Insect Cells 在Sf9昆虫细胞中表达纯化哺乳动物P2X7受体蛋白 作者:Akira Karasawa and Toshimitsu Kawate日期:09/05/2017,浏览量:9009,Q&A: 0
The P2X7 receptor is an extracellular ATP-gated ion channel found only in eukaryotes (Bartlett et al., 2014). Due to its unique properties among P2X receptors, such as formation of a large conductance pore, the P2X7 receptor has been implicated in devastating diseases like chronic pain (North and Jarvis, 2013). However, mechanisms ...
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Detection of Reactive Oxygen Species (ROS) in Cyanobacteria Using the Oxidant-sensing Probe 2’,7’-Dichlorodihydrofluorescein Diacetate (DCFH-DA) 使用氧化物探针2',7'-二氯二氟荧光素二乙酸酯(DCFH-DA)检测蓝细菌中活性氧物质(ROS) 作者:Rajneesh, Jainendra Pathak, Ananya Chatterjee, Shailendra P. Singh and Rajeshwar P. Sinha日期:09/05/2017,浏览量:17357,Q&A: 0
Reactive oxygen species (ROS) are cell signaling molecules synthesized inside the cells as a response to routine metabolic processes. In stress conditions such as ultraviolet radiation (UVR), ROS concentration increases several folds in the cells that become toxic for the cell survival. Here we present the method for in vivo detection of ...
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分子生物学

Using CRISPR-ERA Webserver for sgRNA Design 使用网络服务器CRISPR-ERA设计sgRNA 作者:Honglei Liu, Xiaowo Wang and Lei S. Qi日期:09/05/2017,浏览量:9122,Q&A: 0
The CRISPR-Cas9 system is emerging as a powerful technology for gene editing (modifying the genome sequence) and gene regulation (without modifying the genome sequence). Designing sgRNAs for specific genes or regions of interest is indispensable to CRISPR-based applications. CRISPR-ERA (http://crispr-era.stanford.edu/ ...
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神经科学

FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction 果蝇神经肌肉接头处FM1-43光转换和电子显微镜分析 作者:Nadezhda S. Sabeva and Maria Bykhovskaia日期:09/05/2017,浏览量:6515,Q&A: 0
We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...
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Arginine-rich Peptides Can Actively Mediate Liquid-liquid Phase Separation 富含精氨酸的肽可以主动介导液-液相分离 作者:Steven Boeynaems, Mathias De Decker, Peter Tompa and Ludo Van Den Bosch日期:09/05/2017,浏览量:10495,Q&A: 0
Studying liquid-liquid phase separation (LLPS) of proteins provides key insights into the biogenesis of membraneless organelles and pathological protein aggregation in disease. We have established a protocol for inducing the phase separation of arginine-rich peptides, which allows for studying their molecular determinants and dynamics (Boeynaems ...
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Predator Odor-induced Freezing Test for Mice 捕食者气味诱导的小鼠僵住试验 作者:Shintaro Otsuka日期:09/05/2017,浏览量:6548,Q&A: 0
The innate fear response is an emotional response that does not require any previously acquired conditioning. One of the standard methods to analyze the innate fear response is a 2,4,5-trimethylthiazoline (TMT)-induced freezing test. TMT is an odor originally isolated from anal secretion of the red fox. Acute TMT exposure has been shown to induce ...
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Isolation of Rodent Brain Vessels 啮齿动物脑血管分离 作者:Cristina Carvalho and Paula I. Moreira日期:09/05/2017,浏览量:6961,Q&A: 0
The prevalence of neurodegenerative diseases is increasing worldwide. Cerebrovascular disorders and/or conditions known to affect brain vasculature, such as diabetes, are well-known risk factors for neurodegenerative diseases. Thus, the evaluation of the brain vasculature is of great importance to better understand the mechanisms underlying brain ...
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植物科学

Ubiquitin Proteasome Activity Measurement in Total Plant Extracts 植物总提取物中泛素蛋白酶体活性测定 作者:Suayib Üstün and Frederik Börnke日期:09/05/2017,浏览量:6844,Q&A: 0
The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to ...
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Isolation and Quantification of Plant Extracellular Vesicles 植物细胞外囊泡的分离和定量 作者:Brian D. Rutter, Katie L. Rutter and Roger W. Innes日期:09/05/2017,浏览量:16415,Q&A: 4
Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...
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Extraction and Molybdenum Blue-based Quantification of Total Phosphate and Polyphosphate in Parachlorella 拟小球藻中总磷酸盐和多磷酸盐的提取和钼蓝法定量 作者:Shuhei Ota and Shigeyuki Kawano日期:09/05/2017,浏览量:8167,Q&A: 0
Inorganic phosphorus is a non-renewable resource and an essential element for life on Earth. Organisms such as algae, protists, and animals can store phosphate (Pi) through uptake of Pi as polyphosphate (poly-P), which is a linear polymer of orthophosphate residues linked by high-energy phosphoanhydride bonds. Here, we describe procedures for ...
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Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants 构建共表达Cas9和sgRNAs的单一独立转录单元用于植物基因组编辑 作者:Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang日期:09/05/2017,浏览量:10220,Q&A: 0
The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are typically expressed from RNA polymerase III promoters, such as U6 and U3. In many transformation events, ...
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