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Image Credit: BioProtoc.2990

生物化学

Structural Analysis of Target Protein by Substituted Cysteine Accessibility Method 采用半胱氨酸取代法分析靶蛋白的结构 作者:Tetsuo Cai and Taisuke Tomita日期:09/05/2018,浏览量:4544,Q&A: 0
Substituted Cysteine Accessibility Method (SCAM) is a biochemical approach to investigate the water accessibility or the spatial distance of particular cysteine residues substituted in the target protein. Protein topology and structure can be annotated by labeling with methanethiosulfonate reagents that specifically react with the cysteine ...
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Detection of Internal Matrix Targeting Signal-like Sequences (iMTS-Ls) in Mitochondrial Precursor Proteins Using the TargetP Prediction Tool 用TargetP预测工具在线粒体前体蛋白中检测内部类基质导向信号序列 作者:Felix Boos, Timo Mühlhaus and Johannes M. Herrmann日期:09/05/2018,浏览量:5598,Q&A: 0
Mitochondria contain hundreds of proteins which are encoded by the nuclear genome and synthesized in the cytosol from where they are imported into the organelle. Sorting signals encoded in the primary and secondary sequence of these proteins mediate the recognition of newly synthesized precursor proteins and their subsequent translocation through ...
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癌症生物学

Cell Synchronization by Double Thymidine Block 用胸腺嘧啶核苷双阻断法同步细胞 作者:Guo Chen and Xingming Deng日期:09/05/2018,浏览量:25647,Q&A: 1
Cell synchronization is widely used in studying mechanisms involves in regulation of cell cycle progression. Through synchronization, cells at distinct cell cycle stage could be obtained. Thymidine is a DNA synthesis inhibitor that can arrest cell at G1/S boundary, prior to DNA replication. Here, we present the protocol to synchronize cells at ...
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发育生物学

Human Endothelial Cell Spheroid-based Sprouting Angiogenesis Assay in Collagen 在胶原蛋白中分析人内皮细胞基于球体的出芽式血管生成 作者:Fabian Tetzlaff and Andreas Fischer日期:09/05/2018,浏览量:13505,Q&A: 0
Angiogenesis, the formation of new blood vessels from pre-existing ones plays an important role during organ development, regeneration and tumor progression. The spheroid-based sprouting assay is a well-established and robust method to study the influence of genetic alterations or pharmacological compounds on capillary-like tube formation of ...
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免疫学

Using Stable Isotopes in Bone Marrow Derived Macrophage to Analyze Metabolism 利用稳定同位素研究骨髓源性巨噬细胞的代谢 作者:Chih-Wei Ko, Daniel Counihan, David DeSantis, Zach Sedor-Schiffhauer, Michelle Puchowicz and Colleen M Croniger日期:09/05/2018,浏览量:5292,Q&A: 0
Using gas chromatography mass spectrometry (GC-MS) to analyze the citric acid cycle (CAC) and related intermediates (such as glutamate, glutamine, GABA, and aspartate) is an analytical approach to identify unexpected correlations between apparently related and unrelated pathways of energy metabolism. Intermediates can be as expressed as their ...
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微生物学

Artificial Inoculation of Epichloë festucae into Lolium perenne, and Visualisation of Endophytic and Epiphyllous Fungal Growth 人工接种羊茅香柱菌到麦黑草,并可视化观察内生真菌和叶生真菌的生长 作者:Yvonne Becker, Kimberly A. Green, Barry Scott and Matthias Becker日期:09/05/2018,浏览量:6340,Q&A: 0
Natural hosts for the fungal endophyte Epichloë festucae include Festuca rubra (fine fescue) and Festuca trachyphylla (hard fescue). Some strains also form stable associations with Lolium perenne (perennial ryegrass). L. perenne is a suitable host to study fungal endophyte–grass interactions, such as endophytic ...
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Analysis of the Effect of Sphingomyelinase on Rubella Virus Infectivity in Two Cell Lines 在两细胞系中分析鞘磷脂酶对于风疹病毒感染力的影响 作者:Noriyuki Otsuki, Masafumi Sakata, Yoshio Mori, Kiyoko Okamoto and Makoto Takeda日期:09/05/2018,浏览量:4107,Q&A: 0
Rubella is a mildly contagious disease characterized by low-grade fever and a morbilliform rash caused by the rubella virus (RuV). Viruses often use cellular phospholipids for infection. We studied the roles of cellular sphingomyelin in RuV infection. Treatment of cells with sphingomyelinase (SMase) inhibited RuV infection in rabbit kidney-derived ...
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Soluble and Solid Iron Reduction Assays with Desulfitobacterium hafniense 哈夫尼脱亚硫酸杆菌用于可溶和固体铁的还原试验 作者:Lucrezia Comensoli, Julien Maillard, Wafa M. Kooli, Pilar Junier and Edith Joseph日期:09/05/2018,浏览量:4375,Q&A: 0
There is a pressing need to develop sustainable and efficient methods to protect and stabilize iron objects. To develop a conservation-restoration method for corroded iron objects, this bio-protocol presents the steps to investigate reductive dissolution of ferric iron and biogenic production of stabilizing ferrous iron minerals in the strict ...
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Selective Isolation of Retroviruses from Extracellular Vesicles by Intact Virion Immunoprecipitation 通过完整病毒免疫沉淀技术从细胞外囊泡选择性分离逆转录病毒 作者:Tyler Milston Renner, Kasandra Bélanger and Marc-André Langlois日期:09/05/2018,浏览量:9482,Q&A: 0
There exists a wide variety of techniques to isolate and purify viral particles from cell culture supernatants. However, these techniques vary greatly in ease of use, purity, yield and impact on viral structural integrity. Most importantly, it is becoming evident that secreted extracellular vesicles (EVs) co-purify with retroviruses using nearly ...
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分子生物学

A Quantitative Heterokaryon Assay to Measure the Nucleocytoplasmic Shuttling of Proteins 定量异核体分析测定蛋白质核质穿梭 作者:François McNicoll and Michaela Müller-McNicoll日期:09/05/2018,浏览量:6742,Q&A: 0
Many proteins appear exclusively nuclear at steady-state but in fact shuttle continuously back and forth between the nucleus and the cytoplasm. For example, nuclear RNA-binding proteins (RBPs) often accompany mRNAs to the cytoplasm, where they can regulate subcellular localization, translation and/or decay of their cargos before shuttling back to ...
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Enzymatic Synthesis and Fractionation of Fluorescent PolyU RNAs 荧光polyU RNAs的酶促合成及分离 作者:Christian Beren, Katherine N. Liu, Lisa L. Dreesens, Charles M. Knobler and William M. Gelbart日期:09/05/2018,浏览量:3335,Q&A: 0
The physical properties of viral-length polyuridine (PolyU) RNAs, which cannot base-pair and form secondary structures, are compared with those of normal-composition RNAs, composed of comparable numbers of each of A, U, G and C nucleobases. In this protocol, we describe how to synthesize fluorescent polyU RNAs using the enzyme polynucleotide ...
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Dual Fluorescence Reporter Based Analytical Flow Cytometry for miRNA Induced Regulation in Mammalian Cells 哺乳动物细胞中利用基于分析流式细胞术的双荧光报告因子研究miRNA诱导调控 作者:Nicolas Lemus-Diaz, Liezel Tamon and Jens Gruber日期:09/05/2018,浏览量:9063,Q&A: 0
MicroRNA-induced gene regulation is a growing field in basic and translational research. Examining this regulation directly in cells is necessary to validate high-throughput data originated from RNA sequencing technologies. For this several studies employ luciferase-based reporters that usually measure the whole cell population, which comes with ...
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神经科学

Phagocytosis Assay for α-Synuclein Fibril Uptake by Mouse Primary Microglia 小鼠初级小胶质细胞吸收α-突触核蛋白原纤维的吞噬分析 作者:Cuilian Du, Feifan Zhang and Claire Xi Zhang日期:09/05/2018,浏览量:4360,Q&A: 0
Microglia are professional phagocytes in the brain and deficiency in their phagocytic activity plays an important role in Parkinson’s disease. This protocol mainly describes the phagocytosis assay for uptake of α-synuclein preformed fibrils, a pathologic form of α-synuclein, by primary microglia.
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Pentylenetetrazole (PTZ)-induced Convulsion Assay to Determine GABAergic Defects in Caenorhabditis elegans 用于确定秀丽隐杆线虫γ-氨基丁酸能缺陷的戊四氮唑诱导的惊厥测定试验 作者:Shruti Thapliyal and Kavita Babu日期:09/05/2018,浏览量:4074,Q&A: 0
Pentylenetetrazole (PTZ) is a GABAA receptor antagonist and is used to monitor presynaptic defects in the release of the inhibitory neurotransmitter GABA. PTZ is a competitive inhibitor of GABA, and prevents binding of GABA on the GABAA receptors present on the surface of muscle. In the absence of GABA binding, the excitatory ...
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Shock-probe Defensive Burying Test to Measure Active versus Passive Coping Style in Response to an Aversive Stimulus in Rats 冲击探测防御性埋藏实验测定大鼠响应厌恶性刺激的积极或消极应对方式 作者:Elizabeth A. Fucich and David A. Morilak日期:09/05/2018,浏览量:4599,Q&A: 0
Maladaptive avoidance behaviors are seen in many stress-related psychiatric illnesses. Patients with these illnesses favor passive, avoidant coping strategies rather than adaptive, active coping strategies. Preclinically, coping strategy can be measured in rats using the shock-probe defensive burying test, wherein rats receive a shock from an ...
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Studying the Mechanisms of Developmental Vocal Learning and Adult Vocal Performance in Zebra Finches through Lentiviral Injection 通过慢病毒注射研究斑马雀的发育声乐学习和成年声乐表演的机制 作者:Zhimin Shi, Ofer Tchernichovski and XiaoChing Li日期:09/05/2018,浏览量:4263,Q&A: 0
Here we provide a detailed step-by-step protocol for using lentivirus to manipulate miRNA expression in Area X of juvenile zebra finches and for analyzing the consequences on song learning and song performance. This protocol has four parts: 1) making the lentiviral construct to overexpress miRNA miR-9; 2) packaging the lentiviral vector; 3) ...
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植物科学

In planta Transcriptome Analysis of Pseudomonas syringae 丁香假单胞菌的植物转录组分析 作者:Tatsuya Nobori and Kenichi Tsuda日期:09/05/2018,浏览量:6643,Q&A: 0
Profiling bacterial transcriptome in planta is challenging due to the low abundance of bacterial RNA in infected plant tissues. Here, we describe a protocol to profile transcriptome of a foliar bacterial pathogen, Pseudomonas syringae pv. tomato DC3000, in the leaves of Arabidopsis thaliana at an early stage of ...
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Enzymatic Assays and Enzyme Histochemistry of Tuta absoluta Feeding on Tomato Leaves 对喂食番茄叶片的番茄潜叶蛾进行酶学测定和酶组织化学检测 作者:Rim Hamza, José P. Beltrán and Luis A. Cañas日期:09/05/2018,浏览量:5312,Q&A: 0
Enzymes play a key role in insect-plant relationships. For a better understanding of these interactions, we analyzed Tuta absoluta digestive enzymes. Here, we describe a detailed protocol for the detection of trypsin and papain-like enzymes in Tuta absoluta larvae by enzyme histochemistry. This assay uses frozen and unfixed ...
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干细胞

Investigating Neural Stem Cell and Glioma Stem Cell Self-renewal Potential Using Extreme Limiting Dilution Analysis (ELDA) 利用极端有限稀释分析法(ELDA)检测神经干细胞和胶质瘤干细胞的自我更新能力 作者:Hong PT Nguyen, Paul M Daniel, Gulay Filiz and Theo Mantamadiotis日期:09/05/2018,浏览量:6799,Q&A: 0
Glioma stem cells (GSC) grown as neurospheres exhibit similar characteristics to neural stem cells (NSC) grown as neurospheres, including the ability to self-renew and differentiate. GSCs are thought to play a role in cancer initiation and progression. Self-renewal potential of GSCs is thought to reflect many characteristics associated with ...
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Microarray, IPA and GSEA Analysis in Mice Models 关于小鼠模型的微阵列,IPA和GSEA分析 作者:Stephanie N. Oprescu, Katharine A. Horzmann, Feng Yue, Jennifer L. Freeman and Shihuan Kuang日期:09/05/2018,浏览量:7622,Q&A: 0
This protocol details a method to analyze two tissue samples at the transcriptomic level using microarray analysis, ingenuity pathway analysis (IPA) and gene set enrichment analysis (GSEA). Methods such as these provide insight into the mechanisms underlying biological differences across two samples and thus can be applied to interrogate a variety ...
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