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生物化学

Active Cdk5 Immunoprecipitation and Kinase Assay 活性Cdk5免疫沉淀及其激酶活性的测定 作者:Andrew N. Bankston, Li Ku and Yue Feng日期:07/05/2017,浏览量:7510,Q&A: 0
Cdk5 activity is regulated by the amounts of two activator proteins, p35 and p39 (Tsai et al., 1994; Zheng et al., 1998; Humbert et al., 2000). The p35-Cdk5 and p39-Cdk5 complexes have differing sensitivity to salt and detergent concentrations (Hisanaga and Saito, 2003; Sato et al., 2007; Yamada et al., ...
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Formaldehyde Fixation of Extracellular Matrix Protein Layers for Enhanced Primary Cell Growth 甲醛法固定细胞外基质蛋白层以增强原代细胞生长 作者:Natalia V. Andreeva and Alexander V. Belyavsky日期:07/05/2017,浏览量:7093,Q&A: 0
Coating tissue culture vessels with the components of the extracellular matrix such as fibronectin and collagens provides a more natural environment for primary cells in vitro and stimulates their proliferation. However, the effects of such protein layers are usually rather modest, which might be explained by the loss immobilized proteins ...
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癌症生物学

Tumorigenicity Assay in Nude Mice 裸鼠中的致瘤性试验 作者:Feng Du, Xiaodi Zhao and Daiming Fan日期:07/05/2017,浏览量:17064,Q&A: 1
Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and the tumor growth is monitored over a period of ...
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Generation of a Cellular Reporter for Functional BRD4 Inhibition BRD4抑制功能细胞报告基因的生成 作者:Sara Sdelci and Stefan Kubicek日期:07/05/2017,浏览量:5018,Q&A: 0
The ubiquitously expressed bromodomain-containing protein 4 (BRD4) is an epigenetic reader, which recruits transcriptional regulatory complexes to acetylated chromatin. Because of its role in enhancing proliferation, BRD4 has become a therapeutic target in oncology, as the inhibition of this protein leads to the reduction of the growth of many ...
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A Novel Mouse Skin Graft Model of Vascular Tumors Driven by Akt1 由Akt1驱动的血管肿瘤的新型小鼠皮肤移植模型 作者:Thuy L. Phung and Sriram Ayyaswamy日期:07/05/2017,浏览量:8711,Q&A: 0
To investigate whether endothelial Akt1 activation is sufficient to induce vascular tumor formation in the skin, we have developed a skin graft model in which a skin fragment from transgenic donor mice with inducible and endothelial cell-specific overexpression of activated Akt1 (myrAkt1) is grafted into the skin of wild type recipient mice. The ...
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细胞生物学

Photothrombotic Induction of Capillary Ischemia in the Mouse Cortex during in vivo Two-Photon Imaging 在体内双光子成像期间采用光栓法诱导小鼠皮层毛细血管缺血 作者:Robert G. Underly and Andy Y. Shih日期:07/05/2017,浏览量:7431,Q&A: 0
Photothrombosis of blood vessels refers to the activation of a circulating photosensitive dye with a green light to induce clotting in vivo (Watson et al., 1985). Previous studies have described how a focused green laser could be used to noninvasively occlude pial arterioles and venules at the brain surface (Schaffer et al. ...
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Endpoint or Kinetic Measurement of Hydrogen Sulfide Production Capacity in Tissue Extracts 组织提取物中生成硫化氢的终点或动力学测定 作者:Christopher Hine and James R. Mitchell日期:07/05/2017,浏览量:8092,Q&A: 0
Hydrogen sulfide (H2S) gas is produced in cells and tissues via various enzymatic processes. H2S is an important signaling molecule in numerous biological processes, and deficiencies in endogenous H2S production are linked to cardiovascular and other health complications. Quantitation of steady-state H2S ...
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发育生物学

Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans 秀丽隐杆线虫中氧化应激指标(亚砷酸盐和tBHP)的测定 作者:Collin Yvès Ewald, John M. Hourihan and T. Keith Blackwell日期:07/05/2017,浏览量:9610,Q&A: 0
Cells and organisms face constant exposure to reactive oxygen species (ROS), either from the environment or as a by-product from internal metabolic processes. To prevent cellular damage from ROS, cells have evolved detoxification mechanisms. The activation of these detoxification mechanisms and their downstream responses represent an overlapping ...
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Ex vivo Ooplasmic Extract from Developing Drosophila Oocytes for Quantitative TIRF Microscopy Analysis 从在发育的果蝇卵母细胞中离体提取卵磷脂用于定量TIRF显微镜分析 作者:Imre Gáspár and Anne Ephrussi日期:07/05/2017,浏览量:6511,Q&A: 0
Understanding the dynamic behavior and the continuously changing composition of macromolecular complexes, subcellular structures and organelles is one of areas of active research in both cell and developmental biology, as these changes directly relate to function and subsequently to the development and homeostasis of the organism. Here, we ...
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免疫学

Thermal Stability of Heterotrimeric pMHC Proteins as Determined by Circular Dichroism Spectroscopy 采用圆二色光谱法测定异源三聚体pMHC蛋白质的热稳定性 作者:Anna Fuller, Aaron Wall, Michael D Crowther, Angharad Lloyd, Alexei Zhurov, Andrew K. Sewell, David K. Cole and Konrad Beck日期:07/05/2017,浏览量:6699,Q&A: 0
T cell receptor (TCR) recognition of foreign peptide fragments, presented by peptide major histocompatibility complex (pMHC), governs T-cell mediated protection against pathogens and cancer. Many factors govern T-cell sensitivity, including the affinity of the TCR-pMHC interaction and the stability of pMHC on the surface of antigen presenting ...
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Qualitative and Quantitative Assay for Detection of Circulating Autoantibodies against Human Aortic Antigen 人主动脉抗原循环自身抗体的定性和定量检测实验 作者:Brent Veerman and Ritu Chakravarti日期:07/05/2017,浏览量:7913,Q&A: 0
Increased amount of autoantibodies in human sera are the hallmark of autoimmune diseases (Wang et al., 2015). In case of known antigen, detection of autoantibodies is done using laboratory based methods. However, in most autoimmune diseases, knowledge of self-antigen is still vague. We have developed an ELISA-based quantitative assay to ...
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EAE Induction by Passive Transfer of MOG-specific CD4+ T Cells 通过MOG特异性CD4+ T细胞的被动转移诱导EAE 作者:Yuki Tanaka, Yasunobu Arima, Kotaro Higuchi, Takuto Ohki, Mohamed Elfeky, Mitsutoshi Ota, Daisuke Kamimura and Masaaki Murakami日期:07/05/2017,浏览量:9299,Q&A: 0
Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which is a chronic inflammatory disease of the central nervous system (CNS). It is characterized by focal demyelination and inflammatory responses mediated by myelin-specific autoreactive CD4+ T cells. Using a passive transfer model of EAE in ...
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Vaginal HSV-2 Infection and Tissue Analysis 阴道HSV-2感染和组织分析 作者:Marie Beck Iversen, Søren Riis Paludan and Christian Kanstrup Holm日期:07/05/2017,浏览量:7729,Q&A: 0
The vaginal murine HSV-2 infection model is very useful in studying mucosal immunity against HSV-2 (Overall et al., 1975; Renis et al., 1976; Parr and Parr, 2003). Histologically, the vagina of Depo-Provera-treated mice is lined by a single layer of mucus secreting columnar epithelial cells overlying two to three layers of ...
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In vitro Demonstration and Quantification of Neutrophil Extracellular Trap Formation 中性粒细胞胞外诱捕网形成的体外演示和定量 作者:Dongsheng Jiang, Mona Saffarzadeh and Karin Scharffetter-Kochanek日期:07/05/2017,浏览量:12479,Q&A: 0
In the recent decade, neutrophil extracellular traps (NETs) have been identified and confirmed as a new anti-microbial weapon of neutrophils. In this protocol, we describe easy methods to demonstrate NET formation by immunofluorescence staining of extracellular chromatin fiber with anti-DNA/Histone H1 antibody and quantification of NETs by using a ...
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微生物学

Assaying Glycogen and Trehalose in Yeast 酵母中糖原和海藻糖的测定 作者:Yuping Chen and Bruce Futcher日期:07/05/2017,浏览量:8081,Q&A: 0
Organisms store carbohydrates in several forms. In yeast, carbohydrates are stored in glycogen (a multi-branched polysaccharide) and in trehalose (a disaccharide). As in other organisms, the amount of stored carbohydrate varies dramatically with physiological state, and accordingly, an assay of stored carbohydrate can help reveal physiological ...
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Bacterial Survival in Dictyostelium 盘基网柄菌中细菌存活能力的检测 作者:Regin Rønn, Xiuli Hao, Freja Lüthje, Nadezhda A. German, Xuanji Li, Fuyi Huang, Javan Kisaka, David Huffman, Hend A. Alwathnani, Yong-Guan Zhu and Christopher Rensing日期:07/05/2017,浏览量:6573,Q&A: 0
We performed an assay to test the ability of different E. coli strains to survive inside amoebal cells after ingestion. In the assay we incubated bacteria together with cells of Dictyostelium discoideum for six hours. After co-incubation most of the uningested bacteria were removed by centrifugation and the remaining uningested ...
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Determination of NO and CSF Levels Produced by Bacillus subtilis 枯草芽孢杆菌生成的NO和CSF水平测定 作者:Sebastián Cogliati, Facundo Rodriguez Ayala, Carlos Bauman, Marco Bartolini, Cecilia Leñini, Juan Manuel Villalba, Federico Argañaraz and Roberto Grau日期:07/05/2017,浏览量:6584,Q&A: 0
The cell-to-cell communication and division of labour that occurs inside a beneficial biofilm produce significant differences in gene expression compared with the gene expression pattern of cells grew under planktonic conditions. In this sense, the levels of NO (nitric oxide) and CSF (Competence Sporulation Stimulating Factor) produced in Bacillus ...
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分子生物学

Multiplex Gene Editing via CRISPR/Cas9 System in Sheep 通过CRISPR/Cas9系统在绵羊中进行多重基因编辑 作者:Yiyuan Niu, Yi Ding, Xiaolong Wang and Yulin Chen日期:07/05/2017,浏览量:8537,Q&A: 0
Sheep is a major large animal model for studying development and disease in biomedical research. We utilized CRISPR/Cas9 system successfully to modify multiple genes in sheep. Here we provide a detailed protocol for one-cell-stage embryo manipulation by co-injecting Cas9 mRNA and RNA guides targeting three genes (MSTN, ASIP, and ...
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神经科学

Social Observation Task in a Linear Maze for Rats 大鼠线性迷宫社会观察任务实验 作者:Xiang Mou and Daoyun Ji日期:07/05/2017,浏览量:5499,Q&A: 0
Animals often learn through observing their conspecifics. However, the mechanisms of them obtaining useful knowledge during observation are beginning to be understood. This protocol describes a novel social observation task to test the ‘local enhancement theory’, which proposes that presence of social subjects in an environment facilitates one’s ...
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Live Imaging of Myogenesis in Indirect Flight Muscles in Drosophila 果蝇间接飞行肌中肌生成的实时成像 作者:Dagan Segal日期:07/05/2017,浏览量:7354,Q&A: 0
The indirect flight muscles (IFMs) are the largest muscles in the fly, making up the bulk of the adult thorax. IFMs in Drosophila are generated during pupariation by fusion of hundreds of muscle precursor cells (myoblasts) with larval muscle templates (myotubes). Prominent features, including the large number of fusion events, the ...
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Spinal Cord Preparation from Adult Red-eared Turtles for Electrophysiological Recordings during Motor Activity 制备成年红耳龟脊髓以用于运动活动期间电生理的记录 作者:Peter C Petersen and Rune W Berg 日期:07/05/2017,浏览量:9779,Q&A: 0
Although it is known that the generation of movements is performed to a large extent in neuronal circuits located in the spinal cord, the involved mechanisms are still unclear. The turtle as a model system for investigating spinal motor activity has advantages, which far exceeds those of model systems using other animals. The high resistance to ...
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Behavioral and Functional Assays for Investigating Mechanisms of Noxious Cold Detection and Multimodal Sensory Processing in Drosophila Larvae 行为和功能分析实验研究蝇幼虫感知有害寒冷及处理多模态感知的机制 作者:Atit A. Patel and Daniel N. Cox日期:07/05/2017,浏览量:7346,Q&A: 0
To investigate cellular, molecular and behavioral mechanisms of noxious cold detection, we developed cold plate behavioral assays and quantitative means for evaluating the predominant noxious cold-evoked contraction behavior. To characterize neural activity in response to noxious cold, we implemented a GCaMP6-based calcium imaging assay enabling ...
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Delayed-matching-to-place Task in a Dry Maze to Measure Spatial Working Memory in Mice 采用日间迷宫中延迟位置匹配任务来衡量小鼠的空间工作记忆 作者:Xi Feng, Karen Krukowski, Timothy Jopson and Susanna Rosi日期:07/05/2017,浏览量:6970,Q&A: 0
The delayed-matching-to-place (DMP) dry maze test is a variant of DMP water maze (Steele and Morris, 1999; Faizi et al., 2012) which measures spatial working/episodic-like learning and memory that depends on both hippocampal and cortical functions (Wang and Morris, 2010; Euston et al., 2012). Using this test we can detect normal ...
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植物科学

A Simple and Rapid Assay for Measuring Phytoalexin Pisatin, an Indicator of Plant Defense Response in Pea (Pisum sativum L.) 一种简单快速测定豌豆(Pisum sativum L.)中植物防御反应指标-植物抗毒素豌豆素的方法 作者:Lee A. Hadwiger and Kiwamu Tanaka日期:07/05/2017,浏览量:5972,Q&A: 0
Phytoalexins are antimicrobial substance synthesized in plants upon pathogen infection. Pisatin (Pisum sativum phytoalexin) is the major phytoalexin in pea, while it is also a valuable indicator of plant defense response. Pisatin can be quantitated in various methods from classical organic chemistry to ...
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Determination of Reduced and Total Glutathione Content in Extremophilic Microalga Galdieria phlegrea 极端环境微藻Galdieria phlegrea中还原型和总谷胱甘肽含量的测定 作者:Giovanna Salbitani, Claudia Bottone and Simona Carfagna日期:07/05/2017,浏览量:17137,Q&A: 3
Glutathione is an important molecule involved in the primary and secondary metabolism of all organisms. The Glutathione redox status is an indicator of the cellular redox state. Therefore, it is important to have precise methods on hand to determine the glutathione redox status in the cell. In this protocol, we describe an improved ...
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Generation of Targeted Knockout Mutants in Arabidopsis thaliana Using CRISPR/Cas9 拟南芥中利用CRISPR/Cas9技术生成定向基因敲除突变体 作者:Florian Hahn, Marion Eisenhut, Otho Mantegazza and Andreas P. M. Weber日期:07/05/2017,浏览量:19696,Q&A: 1
The CRISPR/Cas9 system has emerged as a powerful tool for gene editing in plants and beyond. We have developed a plant vector system for targeted Cas9-dependent mutagenesis of genes in up to two different target sites in Arabidopsis thaliana. This protocol describes a simple 1-week cloning procedure for a single T-DNA vector containing ...
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Determination of the Effects of Local and Systemic Iron Excess on Lateral Root Initiation in Arabidopsis thaliana 局部和系统性铁过量对拟南芥侧根起始影响的测定 作者:Guangjie Li, Lin Zhang and Weiming Shi日期:07/05/2017,浏览量:7797,Q&A: 0
Root system architecture depends on nutrient availability. A symptom of iron (Fe) toxicity in plants is stunted root growth, yet little is known about the effects of excess Fe on lateral root (LR) development. To better understand how nutrient signals are integrated into root developmental programs, we investigated the morphological response of ...
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干细胞

Microvesicle Isolation from Rat Brain Extract Treated Human Mesenchymal Stem Cells 从大鼠脑提取物处理的人间充质干细胞中分离微泡 作者:Ji Yong Lee, Seong-Mi Choi and Han-Soo Kim日期:07/05/2017,浏览量:8642,Q&A: 0
Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their ...
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