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Ca2+ Measurements in Mammalian Cells with Aequorin-based Probes 使用基于水母发光蛋白的探针测定哺乳动物细胞中的Ca2+ 作者:Anna Tosatto, Rosario Rizzuto and Cristina Mammucari日期:03/05/2017,浏览量:6172,Q&A: 0
Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca2+ uptake and release of different ...
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Liposome Flotation Assays for Phosphoinositide-protein Interaction 磷酸肌醇-蛋白质相互作用的脂质体浮选分析法 作者:Helene Tronchere and Frederic Boal日期:03/05/2017,浏览量:15779,Q&A: 0
Phosphoinositides are rare membrane lipids involved in the control of the major cellular functions and signaling pathways. They are able to recruit specific effector proteins to the cytosolic face of plasma membrane and organelles to coordinate a vast variety of signaling and trafficking processes, as well to maintain specific identity of the ...
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Protein Synthesis Rate Assessment by Fluorescence Recovery after Photobleaching (FRAP) 采用荧光漂白恢复法(FRAP)评估蛋白质合成速率 作者:Nikos Kourtis and Nektarios Tavernarakis日期:03/05/2017,浏览量:7483,Q&A: 0
Currently available biochemical methods cannot be applied to monitor protein synthesis in specific cells or tissues, in live specimens. Here, we describe a non-invasive method for monitoring protein synthesis in single cells or tissues with intrinsically different translation rates, in live Caenorhabditis elegans animals.
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An HPLC-based Method to Quantify Coronatine Production by Bacteria 采用基于HPLC的方法定量细菌的冠菌素生成 作者:Shweta Panchal, Zachary S. Breitbach and Maeli Melotto日期:03/05/2017,浏览量:7034,Q&A: 0
Coronatine is a polyketide phytotoxin produced by several pathovars of the plant pathogenic bacterium Pseudomonas syringae. It is one of the most important virulence factors determining the success of bacterial pathogenesis in the plant at both epiphytic and endophytic stages of the disease cycle. This protocol describes an optimized ...
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Polyethylene Glycol-mediated Transformation of Drechmeria coniospora 聚乙二醇介导的圆锥掘氏梅里霉的转化 作者:Le D. He and Jonathan J. Ewbank日期:03/05/2017,浏览量:7184,Q&A: 0
Drechmeria coniospora is a nematophagous fungus and potential biocontrol agent. It belongs to the Ascomycota. It is related to Hirsutella minnesotensis, another nematophagous fungus but, phylogenetically, it is currently closest to the truffle parasite Tolypocladium ophioglossoides. Together with its natural host, ...
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Next-generation Sequencing of the DNA Virome from Fecal Samples 粪便样本中DNA病毒组的下一代测序法 作者:Cynthia L. Monaco and Douglas S. Kwon日期:03/05/2017,浏览量:8055,Q&A: 0
Herein we describe a detailed protocol for DNA virome analysis of low input human stool samples (Monaco et al., 2016). This protocol is divided into four main steps: 1) stool samples are pulverized to evenly distribute microbial matter; 2) stool is enriched for virus-like particles and DNA is extracted by phenol-chloroform; 3) purified ...
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Isolation of Outer Membrane Vesicles from Phytopathogenic Xanthomonas campestris pv. campestris 从野油菜黄单胞菌野油菜致病变种分离外膜囊泡 作者:Gideon Mordukhovich and Ofir Bahar日期:03/05/2017,浏览量:9114,Q&A: 5
Gram-negative bacteria naturally release outer membrane vesicles (OMVs) to the surrounding environment. OMVs contribute to multiple processes, such as cell-cell communication, delivery of enzymes and toxins, resistance to environmental stresses and pathogenesis. Little is known about OMVs produced by plant-pathogenic bacteria, and their ...
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Transient Transfection-based Fusion Assay for Viral Proteins 基于瞬时转染的病毒蛋白融合分析法 作者:Melina Vallbracht, Christina Schröter, Barbara G. Klupp and Thomas C. Mettenleiter日期:03/05/2017,浏览量:9153,Q&A: 1
Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) glycoproteins gB, gH and gL. Here, we describe an ...
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Chitin Extraction and Content Measurement in Magnaporthe oryzae 稻瘟病菌中的甲壳素提取和含量测定 作者:Xinyu Liu and Zhengguang Zhang日期:03/05/2017,浏览量:6713,Q&A: 0
Chitin is a linear polysaccharide composed of β (1→4)-linked N-acetylglucosamine (GlcNAc) residues. In fungi, chitin is an important component of the cell wall. Here, we provide a protocol to measure the chitin content of fungal cells using Magnaporthe oryzae as an example.
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Determination of Elemental Concentrations in Lichens Using ICP-AES/MS ICP-AES/MS法测定地衣中元素的浓度 作者:Liang-Cheng Zhao, Li Wang, Yun-Jun Jiang, Yan-Qiao Hu, Chong-Ying Xu, Lei Wang, Xing Li , Li Wei, Xiu-Ping Guo, Ai-Qin Liu and Hua-Jie Liu日期:03/05/2017,浏览量:8023,Q&A: 1
Lichens are good biomonitors for air pollution because of their high enrichment capability of atmospheric chemical elements. To monitor atmospheric element deposition using lichens, it is important to obtain information on the multi-element concentrations in lichen thalli. Because of serious air pollution, elemental concentrations in thalli of ...
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Extracellular Axon Stimulation 细胞外轴突刺激 作者:Carola Städele, Margaret Louise DeMaegd and Wolfgang Stein日期:03/05/2017,浏览量:9879,Q&A: 0
This is a detailed protocol explaining how to perform extracellular axon stimulations as described in Städele and Stein, 2016. The ability to stimulate and record action potentials is essential to electrophysiological examinations of neuronal function. Extracellular stimulation of axons traveling in fiber bundles (nerves) is a classical technique ...
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Axonal Conduction Velocity Measurement 轴向传导速度测量 作者:Margaret Louise DeMaegd, Carola Städele and Wolfgang Stein日期:03/05/2017,浏览量:9325,Q&A: 0
Action potential conduction velocity is the speed at which an action potential (AP) propagates along an axon. Measuring AP conduction velocity is instrumental in determining neuron health, function, and computational capability, as well as in determining short-term dynamics of neuronal communication and AP initiation (Ballo and Bucher, 2009; ...
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Olfactory Avoidance Test (Mouse) 嗅觉回避试验(小鼠) 作者:Hiroo Takahashi and Akio Tsuboi日期:03/05/2017,浏览量:7143,Q&A: 0
In mice, olfaction plays a pivotal role for the various behaviors, such as feeding, mating, nursing and avoidance. Behavioral tests that analyze abilities of odor detection and recognition using genetically modified mice reveal the contribution of target genes to the olfactory processing. Here, we describe the olfactory avoidance test to ...
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Olfactory Habituation-dishabituation Test (Mouse) 嗅觉习惯化-去习惯化试验(小鼠) 作者:Hiroo Takahashi and Akio Tsuboi日期:03/05/2017,浏览量:7768,Q&A: 0
Olfaction plays a fundamental role in the various behaviors such as feeding, mating, nursing, and avoidance in mice. Behavioral tests that characterize abilities of odor detection and recognition using genetically modified mice reveal the contribution of target genes to the olfactory processing. Here, we describe the olfactory ...
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Thinned-skulled Cranial Window Preparation (Mice) 薄颅骨窗口的制备(小鼠) 作者:Lifeng Zhang, Bo Liang, Yun Li and Da-Ting Lin日期:03/05/2017,浏览量:6061,Q&A: 0
Imaging structural plasticity or activity of neurons in the brain circuit will facilitate understanding the neural mechanisms underlying animal behavior. Here we describe a modified procedure, the polished and reinforced thinned-skull cranial window preparation, by which we can image dendrites and spines in mouse layer I cortex for weeks (Zhang et ...
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Direct Visualization and Quantification of the Actin Nucleation and Elongation Events in vitro by TIRF Microscopy 通过TIRF显微镜在体外直接观察和定量肌动蛋白的成核和伸长 作者:Yuxiang Jiang and Shanjin Huang日期:03/05/2017,浏览量:7368,Q&A: 0
Total internal reflection fluorescence (TIRF) microscopy is a powerful tool for visualizing the dynamics of actin filaments at single-filament resolution in vitro. Thanks to the development of various fluorescent probes, we can easily monitor all kinds of events associated with actin dynamics, including nucleation, elongation, bundling, ...
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Acetyl Bromide Soluble Lignin (ABSL) Assay for Total Lignin Quantification from Plant Biomass 采用乙酰溴可溶性木质素(ABSL)测定法定量植物生物量中的总木质素 作者:William J. Barnes and Charles T. Anderson日期:03/05/2017,浏览量:12470,Q&A: 0
Lignin is the second most abundant biopolymer on Earth, providing plants with mechanical support in secondary cell walls and defense against abiotic and biotic stresses. However, lignin also acts as a barrier to biomass saccharification for biofuel generation (Carroll and Somerville, 2009; Zhao and Dixon, 2011; Wang et al., 2013). For ...
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Laser Scanning Confocal Microcopy for Arabidopsis Epidermal, Mesophyll, and Vascular Parenchyma Cells 拟南芥表皮、叶肉和血管实质细胞的激光扫描共聚焦显微镜检查 作者:Christian Elowsky, Yashitola Wamboldt and Sally Mackenzie日期:03/05/2017,浏览量:10996,Q&A: 0
Investigation of protein targeting to plastids in plants by confocal laser scanning microscopy (CLSM) can be complicated by numerous sources of artifact, ranging from misinterpretations from in vivo protein over-expression, false fluorescence in cells under stress, and organellar mis-identification. Our studies have focused on the ...
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Knock-in Blunt Ligation Utilizing CRISPR/Cas9 利用CRISPR/Cas9敲入平端连接 作者:Jonathan M. Geisinger and Michele P. Calos日期:03/05/2017,浏览量:8028,Q&A: 0
The incorporation of the CRISPR/Cas9 bacterial immune system into the genetic engineering toolbox has led to the development of several new methods for genome manipulation (Auer et al., 2014; Byrne et al., 2015). We took advantage of the ability of Cas9 to generate blunt-ended double-strand breaks (Jinek et al., 2012) to ...
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Multiplexed GuideRNA-expression to Efficiently Mutagenize Multiple Loci in Arabidopsis by CRISPR-Cas9 多重向导RNA表达通过CRISPR Cas9对拟南芥中的多个基因座进行高效诱变 作者:Julia Schumacher, Kerstin Kaufmann and Wenhao Yan日期:03/05/2017,浏览量:9528,Q&A: 0
Since the discovery of the CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein (Cas) as an efficient tool for genome editing in plants (Li et al., 2013; Shan et al., 2013; Nekrasov et al., 2013), a large variety of applications, such as gene knock-out, knock-in or transcriptional ...
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Surface Inoculation and Quantification of Pseudomonas syringae Population in the Arabidopsis Leaf Apoplast 拟南芥叶质外体中丁香假单胞菌种群的表面接种和定量 作者:Cristián Jacob, Shweta Panchal and Maeli Melotto日期:03/05/2017,浏览量:10523,Q&A: 1
Bacterial pathogens must enter the plant tissue in order to cause a successful infection. Foliar bacterial pathogens that are not able to directly penetrate the plant epidermis rely on wounds or natural openings to internalize leaves. This protocol describes a procedure to estimate the population size of Pseudomonas syringae in the leaf ...
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In Gel Kinase Assay 凝胶内激酶测定法 作者:Gaston A. Pizzio and Pedro L. Rodriguez日期:03/05/2017,浏览量:12465,Q&A: 0
Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol ‘In Gel Kinase Assay’, which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used ‘In Gel Kinase Assay’ protocol to show that the ...
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Isolation and Primary Culture of Adult Human Adipose-derived Stromal/Stem Cells 成人脂肪源性基质/干细胞的分离和原代培养 作者:Robert B. Jones, Amy L. Strong, Jeffrey M. Gimble and Bruce A. Bunnell日期:03/05/2017,浏览量:11492,Q&A: 0
Adipose-derived stromal/stem cells (ASCs) are multipotent cells that can be isolated from adipose tissue. Studies have shown that cells have the capacity to self-renew and differentiate into adipocyte, chondrocyte, myocyte, and osteoblast lineages. Thus, significant interest regarding their use for regenerative purposes to restore aging or damaged ...
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Reprogram Murine Epiblast Stem Cells by Epigenetic Inhibitors 通过表观遗传抑制剂重新编程鼠外胚层干细胞 作者:Hui Zhang and Yali Dou日期:03/05/2017,浏览量:5448,Q&A: 0
Pluripotent stem cells in the naïve state are highly useful in regenerative medicine and tissue engineering. A robust reprogramming of the primed murine Epiblast Stem Cells (EpiSCs) to naïve pluripotency is feasible via chemical-only approach. This protocol described a method to reprogram murine EpiSCs by MM-401 treatment, which blocks histone ...
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