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生物化学

Protein Flotation Assay to Isolate Lipids Rafts from Soft Tissue or Cells 蛋白质浮选分离试验从软组织或细胞中分离脂筏 作者:Catherine Lemaire-Vieille, Jean Gagnon and Jean-Yves Cesbron日期:08/20/2013,浏览量:9695,Q&A: 0
The arrangement in eukaryotic cell membranes of liquid-ordered states surrounded by liquid-disordered phases allows for the existence of organized membrane microdomains called rafts. Rafts play a crucial role in signal-transduction events, in lipid transport and in various internalization processes, and for all these reasons need to be purified ...
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PTEN-lipid Binding Assay PTEN与脂质结合的试验 作者:Sridhar Kavela, Swapnil R. Shinde and Subbareddy Maddika日期:08/20/2013,浏览量:8747,Q&A: 0
The lipid and protein interactions are an integral and important part of many cellular signaling pathways. The understanding of the selective and specific interaction of the given lipid molecule with the target protein is required for studying cellular signaling. In this assay, different lipids are spotted onto a nitrocellulose membrane to which ...
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癌症生物学

Natural Killer Cell Transfer Assay 自然杀伤细胞迁移试验 作者:Lee-Hwa Tai, Christiano Tanese de Souza, Andrew P. Makrigiannis and Rebecca Ann C. Auer日期:08/20/2013,浏览量:10169,Q&A: 0
Natural Killer (NK) cells are cytotoxic lymphocytes that constitute a major component of the innate immune system. Immunosurveillance of the host by NK cells for malignant and virally-infected cells results in direct cytotoxicity and the production of cytokines to enhance the immune response. This protocol will describe the adoptive transfer of ...
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Quantitative Methylation Specific PCR (qMSP) 特异检测DNA甲基化修饰的定量PCR(qMSP) 作者:Triantafillos Liloglou and Georgios Nikolaidis日期:08/20/2013,浏览量:27815,Q&A: 0
Detection of low copies of methylated DNA targets in clinical specimens is challenging. The quantitative Methylation-Specific PCR (qMSP) assays were designed to specifically amplify bisulphite-converted methylated DNA target sequences in the presence of an excess of unmethylated counterpart sequences. These qMSP assays are real-time PCR assays ...
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细胞生物学

Cell Surface Protein Biotinylation and Analysis 细胞表面蛋白质的生物素修饰和分析 作者:Anna Tarradas, Elisabet Selga, Helena Riuró, Fabiana S. Scornik, Ramon Brugada and Marcel Vergés日期:08/20/2013,浏览量:27148,Q&A: 0
A great way to specifically isolate and quantify proteins in the cell surface membrane is to take advantage of the biotinylation technique. It consists of labeling cell surface proteins with a biotin reagent before lysing the cells, and isolating these tagged proteins by NeutrAvidin pull-down. Then, the samples are subjected to SDS-PAGE ...
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Sodium Current Measurements in HEK293 Cells HEK293细胞中的钠电流测定 作者:Helena Riuró, Anna Tarradas, Elisabet Selga, Ramon Brugada, Fabiana Scornik and Guillermo Pérez日期:08/20/2013,浏览量:12139,Q&A: 0
This protocol is used to measure sodium currents from heterologously transfected cells lines, such as HEK293 cells. Standard whole cell patch clamp technique is used to assess ion channel function. This protocol has been used to study cardiac type Nav1.5 sodium channel, and in particular to compare wild-type and mutant channels related ...
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Fluorescence in situ Hybridization to the Polytene Chromosomes of Anopheles Mosquitoes 疟蚊多线染色体的荧光原位杂交 作者:Ai Xia, Ashley Peery, Maryam Kamali, Jiangtao Liang, Maria V. Sharakhova and Igor V. Sharakhov日期:08/20/2013,浏览量:10728,Q&A: 0
Fluorescence in situ hybridization (FISH) is a method that uses a fluorescently labeled DNA probe for mapping the position of a genetic element on chromosomes. A DNA probe is prepared by incorporating Cy-3 or Cy-5 labeled nucleotides into DNA by nick-translation or a random primed labeling method. This protocol was used to map genes ...
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免疫学

Bronchoalveolar Lavage and Lung Tissue Digestion 支气管肺泡灌洗和肺组织消化 作者:Hongwei Han and Steven F. Ziegler日期:08/20/2013,浏览量:30628,Q&A: 0
Bronchoalveolar lavage (BAL) is a simple but valuable and typically performed technique commonly used for studying the pathogenesis of lung diseases such as asthma and COPD. Cell counts can be combined with new methods for examining inflammatory responses, such as ELISA, Flow cytometric analysis, immunohistochemistry, quantitative polymerase chain ...
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Ex vivo Natural Killer Cell Cytotoxicity Assay 自然杀伤细胞的细胞毒性体外测定 作者:Lee-Hwa Tai, Christiano Tanese de Souza, Andrew P. Makrigiannis and Rebecca Ann C. Auer日期:08/20/2013,浏览量:16795,Q&A: 0
Natural Killer (NK) cells are cytotoxic lymphocytes that constitute a major component of the innate immune system. Immunosurveillance of the host by NK cells for malignant and virally-infected cells results in direct cytotoxicity and the production of cytokines to enhance the immune response. This protocol will describe the “gold standard” ...
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微生物学

Zymogram Assay for the Detection of Peptidoglycan Hydrolases in Streptococcus mutans 采用酶谱分析法检测变异链球菌中的肽聚糖水解酶 作者:Delphine Dufour and Céline M. Lévesque日期:08/20/2013,浏览量:12613,Q&A: 1
Peptidoglycan hydrolases or autolysins are enzymes capable of cleaving covalent bonds in bacterial peptidoglycan cell wall layer. They can participate in the cell division process, in the release of turnover products from peptidoglycan during cell growth, and in cell autolysis induced under particular conditions. The protocol for zymogram ...
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Drug Sensitivity Assay of Xanthomonas citri subsp. citri Using REMA Plate Method 采用REMA孔板法进行柑桔溃疡病亚型的药敏试验 作者:Isabel C. Silva and Henrique Ferreira日期:08/20/2013,浏览量:8924,Q&A: 0
Resazurin Microtiter Assay (REMA) is a simple, rapid, reliable, sensitive, safe and cost-effective measurement of cell viability. Resazurin detects cell viability by converting from a nonfluorescent dye to the highly red fluorescent dye resorufin in response to chemical reduction of growth medium resulting from cell growth (Palomino et al. ...
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Metabolic Labeling of Yeast Sphingolipids with Radioactive D-erythro-[4,5-3H]dihydrosphingosine 使用放射性D-赤氏-[4,5-3氢]二氢神经鞘氨醇对酵母鞘脂类进行代谢标记 作者:Takefumi Karashima, Kentaro Kajiwara and Kouichi Funato日期:08/20/2013,浏览量:8489,Q&A: 0
Yeast sphingolipids can be metabolically labeled with exogenously added radioactive precursors. Here we describe a method to label ceramides and complex sphingolipids of Saccharomyces cerevisiae with a radioactive ceramide precursor, D-erythro-[4, 5-3H]dihydrosphingosine. This protocol is used to study the biosynthesis, ...
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Isolation of Radiolabeled Poliovirus Particles from H1 HeLa Cells 从H1 HeLa细胞中分离放射性标记骨髓灰质炎病毒颗粒 作者:Alexsia Richards and William Jackson日期:08/20/2013,浏览量:6696,Q&A: 0
The following protocol describes the isolation of radioactive viral and subviral particles from infected cells. This protocol has been written for isolation of poliovirus particles from H1 HeLa cells. Infection protocol and timing of [35S] methionine labeling and particle collection should be tailored to the virus of interest. ...
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Bioassay to Screen Pathogenesis-associated Genes in Alternaria brassicicola 筛选甘蓝链格孢菌中致病基因的生物学实验 作者:Yangrae Cho日期:08/20/2013,浏览量:6965,Q&A: 0
Alternaria brassicicola is a necrotrophic fungus that causes black spot disease of most plants in the Brassicaceae, including cultivated Brassica species and weedy Arabidopsis species. Since the concept of transformation constructs of linear minimal elements was developed (Cho et al., 2006), we have ...
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分子生物学

Ultra-low Background DNA Cloning System 高效DNA克隆系统 作者:Yukio Nagano and Kenta Goto日期:08/20/2013,浏览量:15233,Q&A: 0
We have developed a method to clone DNA fragments into the E. coli plasmid vectors with almost 100% efficiency (Goto and Nagano, 2013). This method is based on highly efficient yeast-based in vivo cloning, and the subsequent cloning of the constructed plasmids into E. coli. Our method is useful for various applications: ...
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神经科学

Primary Culture of SVZ-derived Progenitors Grown as Neurospheres 室管膜下区(SVZ)源祖细胞生长为神经球的初代培养 作者:Julien Vernerey, Karine Magalon and Pascale Durbec日期:08/20/2013,浏览量:10196,Q&A: 1
SVZ-derived progenitors grown as neurospheres is a well-known model to study neural stem cell and progenitor functions such as proliferation, differentiation/self-renewal, and migration (Durbec and Rougon, 2001). This protocol is for preparing a culture of SVZ-derived progenitors from 8 early postnatal mouse brains (P0 to P3). One week after cell ...
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Retrograde and Anterograde Tracing of Neural Projections 神经突触的逆行和顺行追踪 作者:Takehiro Kudo and Masahiko Watanabe日期:08/20/2013,浏览量:17282,Q&A: 0
Neurons consist of four elements, the soma, dendrite, axon and terminal. They work in concert as the input (soma and dendrite) and output (axon and terminal) parts of neuronal transmission. To function and maintain neuronal activity and metabolisms, proteins and organelles should be transported from soma to terminal via anterograde axonal ...
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植物科学

Reverse Zymmogram Analysis for the Detection of Protease Inhibitor Activity 反向酶谱分析测定蛋白酶抑制剂活性 作者:Lilia Bernal, Florencia García-Campusano, Edgar Nájera and Felipe Cruz-García日期:08/20/2013,浏览量:10114,Q&A: 0
This protocol describes a gel-based procedure to detect protease inhibitor activity. In this method gelatin is used as a substrate for proteolysis and is copolymerized within the polyacrylamide matrix. Protein extracts are fractioned by SDS-PAGE and then the gel is treated with the protease of interest, which degrades gelatin, except in the areas ...
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Maize Embryo Transient Transformation by Particle Bombardment 基因枪法介导玉米胚的瞬时转化 作者:Matilde Jose-Estanyol日期:08/20/2013,浏览量:7596,Q&A: 0
Particle bombardment has been shown to be a useful method to study gene promoter regulatory elements by transient transformation of maize embryos with different constructions of gene promoters fused to a gene reporter. DNA to transfer is coated to high density gold microparticles and introduced into cells when accelerated by a helium pulse. This ...
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Measuring Germination Percentage in Wheat 小麦发芽率的测定 作者:Harish Manmathan and Nora L.V. Lapitan日期:08/20/2013,浏览量:18004,Q&A: 0
Knowledge of the viability of seeds is a prerequisite for establishing the seeding rate in crop production and for germination-related trait evaluation in crop plants. This method explains a simple procedure to establish germination percentage in wheat seeds.
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Rapid Induction of Water Stress in Wheat 小麦中水分胁迫的快速诱导 作者:Harish Manmathan and Nora L.V. Lapitan日期:08/20/2013,浏览量:9785,Q&A: 0
Traditional water stress evaluation studies in wheat are time consuming and can take up to several months to finish. A rapid phenotypic screening for water stress is important for accommodating time-bound water stress studies such as transient gene silencing studies in wheat. This method explains a procedure to induce water stress in young wheat ...
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Determination of Nitrate Uptake and Accumulation Using 15N in Rice Seedlings 15N 标记法测定水稻幼苗中硝酸盐的吸收和积累 作者:Zhong Tang and Guohua Xu日期:08/20/2013,浏览量:7086,Q&A: 0
Nitrogen-15 is a rare stable isotope of nitrogen. This isotope is often used in agricultural research. For example, Nitrogen-15 is used to trace mineral nitrogen compounds and translocate the nitrogen molecule in plants. This protocol is used to determine nitrate uptake and accumulation in rice seedlings by using Nitrogen-15.
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Measurement of Net NO3- Flux in Rice Plants with the SIET System 水稻NO3-吸收量的测定-SITE法(scanning ion-electrode technique) 作者:Zhong Tang and Guohua Xu日期:08/20/2013,浏览量:6800,Q&A: 0
SIET (scanning ion-electrode technique) is a new technique to study the flow rate of the ions and molecules in real time in living biomaterials by using microelectrodes and microsensor. This technique allows non-invasive, simultaneous measurement of fluxes of specific ions at the surface of an intact plant. It has high temporal and spatial ...
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