• 卷册 11, 2021
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Heparan Sulfate Identification and Characterisation: Method I. Heparan Sulfate Identification by NMR Analysis 硫酸乙酰肝素的鉴定和表征:方法I.通过NMR分析鉴定硫酸乙酰肝素 作者:Susan M. Carnachan and Simon F.R. Hinkley日期:04/05/2017,浏览量:6605,Q&A: 0
Heparin and heparan sulfate (HS) may be purified from complex biological matrices and are often isolated in sub-milligram quantities but not unequivocally identified by spectroscopic means. This protocol details a methodology to rapidly assess the gross compositional features and approximate purity of HS by 1H nuclear magnetic ...
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Heparan Sulfate Identification and Characterisation: Method II. Enzymatic Depolymerisation and SAX-HPLC Analysis to Determine Disaccharide Composition 硫酸乙酰肝素的鉴定和表征:方法II.酶解聚合和SAX-HPLC分析确定二糖组成 作者:Susan M. Carnachan and Simon F.R. Hinkley日期:04/05/2017,浏览量:7320,Q&A: 0
Heparan sulfate (HS) is purified from complex matrices and often not fully characterised to validate its assignment. The characterisation of heparins and heparan sulfates through enzymatic depolymerisation and subsequent strong anion-exchange high performance liquid chromatography (SAX-HPLC) analysis and quantitation of the resulting disaccharides ...
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Isolation and Analysis of Proteoglycans and Glycosaminoglycans from Archaeological Bones and Teeth 考古骨和牙齿中蛋白多糖和糖胺聚糖的分离及分析 作者:Yvette May Coulson-Thomas, Vivien Jane Coulson-Thomas and Andrew L. Norton日期:04/05/2017,浏览量:6011,Q&A: 0
We have developed methods for isolating proteoglycans and glycosaminoglycans from archaeological bones and teeth. These methods have been previously published (Coulson-Thomas et al., 2015) and are described here in more detail. In the case of glycosaminoglycans, the method was a previously described method (Nader et al., 1999) ...
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Secretion of Adipsin as an Assay to Measure Flux from the Endoplasmic Reticulum (ER) 脂肪酶分泌测定作为内质网(ER)流量测定法 作者:Alexandria Brumfield, Natasha Chaudhary and Timothy E. McGraw日期:04/05/2017,浏览量:5942,Q&A: 0
In this protocol we describe a quantitative biochemical assay to assess the efficiency of endoplasmic reticulum (ER) to Golgi protein transport in adipocytes (Bruno et al., 2016). The assay takes advantage of the fact that adipocytes secrete various bioactive proteins, known as adipokines. As a measure of ER to Golgi flux we determine the ...
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Nitroxide Labeling of Proteins and the Determination of Paramagnetic Relaxation Derived Distance Restraints for NMR Studies 蛋白质的硝基氧标记和NMR的顺磁弛豫衍生距离约束的测定 作者:Megan Sjodt and Robert T. Clubb日期:04/05/2017,浏览量:9859,Q&A: 0
Site-specific attachment of paramagnetic spin labels to biomolecules causes distance-dependent line-broadening effects, which can be exploited to study the structure and dynamics of these molecules in solution. This protocol describes how to attach nitroxide spin labels to proteins and how to collect and analyze NMR data using these labeled ...
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In vivo Live Imaging of Calcium Waves and Other Cellular Processes during Fertilization in Caenorhabditis elegans 秀丽隐杆线受精期间钙波和其他细胞过程的体内实时成像 作者:Jun Takayama, Masashi Fujita and Shuichi Onami日期:04/05/2017,浏览量:7688,Q&A: 0
Fertilization calcium waves are a conserved trigger for animal development; however, genetic analysis of these waves has been limited due to the difficulty of imaging in vivo fertilization. Here we describe a protocol to image calcium dynamics during in vivo fertilization in the genetic animal model Caenorhabditis elegans ...
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Gliding Assay to Analyze Microtubule-based Motor Protein Dynamics 采用滑动分析法分析基于微管的运动蛋白动力学 作者:Albert Shim, Gin Tezuka, Luke Kupcha and Li Tao日期:04/05/2017,浏览量:11394,Q&A: 0
The purpose of this protocol is to provide an updated method of performing microtubule gliding assays and visualizing it using fluorescence microscopy.
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In vitro Microtubule Bundling Assay under Physiological Conditions 生理条件下的体外微管成束分析 作者:Zach Geisterfer, Gin Tezuka and Li Tao日期:04/05/2017,浏览量:7112,Q&A: 0
Kinesins play a role in organizing the mitotic spindle through the crosslinking of microtubules (MTs), made possible through binding sites at opposite ends of the holoenzyme. Here, we developed a method to test kinesin MT crosslinking action under physiological conditions.
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Isolation of Mononuclear Cell Populations from Ovarian Carcinoma Ascites 从卵巢癌腹水中分离单核细胞群 作者:Christina Wefers, Ghaith Bakdash, Meritxell Moreno Martin, Tjitske Duiveman-de Boer, Ruurd Torensma, Leon F.A.G. Massuger and I. Jolanda M. de Vries日期:04/05/2017,浏览量:8685,Q&A: 0
Ovarian cancer is one of the most fatal tumors in women. Due to a lack of symptoms and adequate screening methods, patients are diagnosed at advanced stages with extensive tumor burden (Jelovac and Armstrong, 2011). Interestingly, ovarian cancer metastasis is generally found within the peritoneal cavity rather than other tissues (Lengyel, 2010; ...
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In vivo Mitophagy Monitoring in Caenorhabditis elegans to Determine Mitochondrial Homeostasis 秀丽隐杆线虫中的体内线粒体自噬监测以确定线粒体稳态 作者:Konstantinos Palikaras and Nektarios Tavernarakis日期:04/05/2017,浏览量:8003,Q&A: 0
Perturbation of mitochondrial function is a major hallmark of several pathological conditions and ageing, underlining the essential role of fine-tuned mitochondrial activity (Lopez-Otin et al., 2013). Mitochondrial selective autophagy, known as mitophagy, mediates the removal of dysfunctional and/or superfluous organelles, preserving ...
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Assessment of Murine Retinal Function by Electroretinography 通过视网膜电图评估小鼠视网膜功能 作者:Gillie Benchorin, Melissa A. Calton, Marielle O. Beaulieu and Douglas Vollrath日期:04/05/2017,浏览量:12794,Q&A: 0
The electroretinogram (ERG) is a sensitive and noninvasive method for testing retinal function. In this protocol, we describe a method for performing ERGs in mice. Contact lenses on the mouse cornea measure the electrical response to a light stimulus of photoreceptors and downstream retinal cells, and the collected data are analyzed to evaluate ...
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In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae 念珠菌菌丝中嗜中性粒细胞诱捕和攻击后细胞壁变化的体外检测 作者:Alex Hopke and Robert T. Wheeler日期:04/05/2017,浏览量:7254,Q&A: 0
In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell ...
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Isolation of Exosomes from Semen for in vitro Uptake and HIV-1 Infection Assays 从精液中分离外来体用于体外摄取和HIV-1感染测定 作者:Marisa N. Madison, Jennifer L. Welch and Chioma M. Okeoma日期:04/05/2017,浏览量:9144,Q&A: 0
Exosomes are membranous extracellular nanovesicles of endocytic origin. Exosomes are known to carry host and pathogen-derived genomic, proteomic, lipidomic cargos and other extraneous molecules. Exosomes are secreted by diverse cell types into the extracellular milieu and are subsequently internalized by recipient neighboring or distal cells. Upon ...
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Screening for Novel Endogenous Inflammatory Stimuli Using the Secreted Embryonic Alkaline Phosphatase NF-κB Reporter Assay 使用分泌型胚胎碱性磷酸酶NF-κB报告基因实验筛选新型内源性炎症刺激物质 作者:Lorena Zuliani-Alvarez, Anna M. Piccinini and Kim S Midwood日期:04/05/2017,浏览量:7749,Q&A: 0
An immune response can be activated by pathogenic stimuli, as well as endogenous danger signals, triggering the activation of pattern recognition receptors and initiating signalling cascades that lead to inflammation. This method uses THP1-BlueTM cells, a human monocytic cell line which contains an embryonic alkaline phosphatase ...
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RNA Strand Displacement Assay for Hepatitis E Virus Helicase 戊型肝炎病毒解旋酶的RNA链置换测定 作者:Vidya P. Nair and Milan Surjit日期:04/05/2017,浏览量:6566,Q&A: 0
The hepatitis E virus (HEV) helicase uses ATP to unwind the RNA duplexes. This is an essential step for viral replication. This protocol aims to measure the double strand RNA unwinding activity of the HEV helicase.
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RNA-dependent RNA Polymerase Assay for Hepatitis E Virus 戊型肝炎病毒依赖于RNA的RNA聚合酶活性测定 作者:Vidya P. Nair, Saumya Anang, Akriti Srivastava and Milan Surjit日期:04/05/2017,浏览量:8621,Q&A: 0
RNA-dependent RNA polymerase (RdRp) is essential for the replication of viral RNA for RNA viruses. It synthesizes the complementary strand of viral genomic RNA, which is used subsequently as a template to generate more copies of viral genome. This assay measures activity of the hepatitis E virus (HEV) RdRp. In contrast to protocols available to ...
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Measurement of the Galactanase Activity of the GanB Galactanase Protein from Bacillus subtilis 枯草芽孢杆菌中GanB半乳聚糖酶活性的测定 作者:Hildegard Watzlawick日期:04/05/2017,浏览量:6540,Q&A: 0
The activity of the endo-β-1,4-galactanase GanB from B. subtilis on the high molecular weight β-1,4-galactan was determined quantitatively by the measurement of the increase of the reducing power or with the dyed substrate Azo-galactan. The generated degradation products were analyzed using thinlayer-chromatography (TLC) or ...
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ARP2/3 Phosphorylation Assay in the Presence of Recombinant Bacterial Effectors 重组细菌效应子存在条件下ARP2/3磷酸化修饰的检测 作者:Céline Michard, Lawrence L. LeClaire and Patricia Doublet日期:04/05/2017,浏览量:6640,Q&A: 0
The Actin-Related Protein 2/3 (ARP2/3) complex is an actin nucleator that generates a branched actin network in mammalian cells. In addition to binding nucleation promoting factors, LeClaire et al. demonstrated that its phosphorylation state is essential key for its activity (LeClaire et al., 2008). In cells, the ARP2/3 complex ...
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Purification of N-coronafacoyl Phytotoxins from Streptomyces scabies 疮痂病链霉菌N-coronafacoyl植物毒素的纯化 作者:Luke Bown and Dawn R. D. Bignell日期:04/05/2017,浏览量:6157,Q&A: 0
This procedure is used for large-scale purification of N-coronafacoyl phytotoxins that are produced by the potato common scab pathogen Streptomyces scabies. The procedure employs organic extraction of S. scabies culture supernatants under alternating basic and acidic conditions in order to preferentially isolate the ...
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Robust Generation of Knock-in Cell Lines Using CRISPR-Cas9 and rAAV-assisted Repair Template Delivery 使用CRISPR-Cas9和rAAV辅助的修复模板递送稳定生成敲入细胞系 作者:Giel Vandemoortele, Delphine De Sutter and Sven Eyckerman日期:04/05/2017,浏览量:18758,Q&A: 0
The programmable Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease 9 (Cas9) technology revolutionized genome editing by providing an efficient way to cut the genome at a desired location (Ledford, 2015). In mammalian cells, DNA lesions trigger the error-prone non-homologous end joining (NHEJ) DNA repair ...
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Efficient Generation of Multi-gene Knockout Cell Lines and Patient-derived Xenografts Using Multi-colored Lenti-CRISPR-Cas9 使用多色Lenti-CRISPR-Cas9高效生成多基因敲除细胞系和患者源性异种移植物 作者:Lena Behrmann, Scott McComb, Júlia Aguadé-Gorgorió, Yun Huang, Mario Hermann, Pawel Pelczar, Adriano Aguzzi, Jean-Pierre Bourquin and Beat C. Bornhauser日期:04/05/2017,浏览量:16308,Q&A: 0
CRISPR-Cas9 based knockout strategies are increasingly used to analyze gene function. However, redundancies and overlapping functions in biological signaling pathways can call for generating multi-gene knockout cells, which remains a relatively laborious process. Here we detail the application of multi-color LentiCRISPR vectors to simultaneously ...
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Measuring Behavioral Individuality in the Acoustic Startle Behavior in Zebrafish 测量斑马鱼声音惊恐行为中的行为个性 作者:Carlos Pantoja, Adam Hoagland, Elizabeth Carroll, David Schoppik and Ehud Y. Isacoff日期:04/05/2017,浏览量:7688,Q&A: 0
The objective of this protocol is to provide a detailed description for the construction and use of a behavioral apparatus, the zBox, for high-throughput behavioral measurements in larval zebrafish (Danio rerio). The zBox is used to measure behavior in multiple individuals simultaneously. Individual fish are housed in wells of multi-well ...
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Testing Depression in Mice: a Chronic Social Defeat Stress Model 小鼠抑郁症检测:慢性社会挫败应激模型 作者:Hee-Dae Kim, Tanessa Call, Samantha Carotenuto, Ross Johnson and Deveroux Ferguson日期:04/05/2017,浏览量:17512,Q&A: 0
A vast challenge within neuropsychiatric research has been the development of animal models that accurately reflect symptoms associated with affective disorders. An ethologically valid model that has been shown to be effective in studying depression is the chronic social defeat stress model. In this model, C57BL/6J mice are subjected to chronic ...
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Resin-embedded Thin-section Immunohistochemistry Coupled with Triple Cellular Counterstaining 树脂包埋薄层免疫组织化学法联合三重细胞复染 作者:William M Palmer, John W Patrick and Yong-Ling Ruan日期:04/05/2017,浏览量:6173,Q&A: 0
This protocol was developed to study protein localisation within the vascular bundles of developing tomato fruit however, it can be applied to any resin embedded plant tissue. The vascular bundle is comprised of many different cells that all have unique properties. The mature sieve elements are enucleated and contain sieve plates that comprise of ...
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Analyses of Root-secreted Acid Phosphatase Activity in Arabidopsis 拟南芥根分泌酸性磷酸酶活性分析 作者:Liangsheng Wang and Dong Liu日期:04/05/2017,浏览量:8962,Q&A: 0
Induction and secretion of acid phosphatase (APase) is a universal adaptive response of higher plants to low-phosphate stress (Tran et al., 2010). The intracellular APases are likely involved in the remobilization and recycling of phosphate (Pi) from intracellular Pi reserves, whereas the extracellular or secreted APases are believed to ...
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Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration 通过农杆菌浸润法使用双生病毒将CRISPR/Cas9组分转入烟草中 作者:Kangquan Yin, Ting Han and Yule Liu日期:04/05/2017,浏览量:11349,Q&A: 0
CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species (Wang et al., 2016). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end ...
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A Co-culture Model for Determining the Target Specificity of the de novo Generated Retinal Ganglion Cells 使用共培养模型测定新生成的视网膜神经节细胞的靶点特异性 作者:Pooja Teotia, Matthew J. Van Hook and Iqbal Ahmad日期:04/05/2017,浏览量:7030,Q&A: 0
In glaucoma, the output neurons of the retina, the retinal ganglion cells (RGCs), progressively degenerate, leading to irreversible blindness (Ahram et al., 2015). The ex vivo stem cell method to replace degenerated RGCs remains a potentially viable approach (Levin et al., 2004). However, the success of the approach ...
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