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Image Credit: BioProtoc.2790

生物化学

Guanine Nucleotide Exchange Assay Using Fluorescent MANT-GDP 使用荧光MANT-GDP的鸟嘌呤核苷酸交换测定法 作者:Tomoharu Kanie and Peter K. Jackson日期:04/05/2018,浏览量:10494,Q&A: 0
GTPases are molecular switches that cycle between the inactive GDP-bound state and the active GTP-bound state. GTPases exchange nucleotides either by its intrinsic nucleotide exchange or by interaction with guanine nucleotide exchange factors (GEFs). Monitoring the nucleotide exchange in vitro, together with reconstitution of direct ...
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生物物理学

Xenopus laevis Oocytes Preparation for in-Cell EPR Spectroscopy 制备非洲爪蟾卵母细胞用于细胞内EPR光谱学分析 作者:Laura John and Malte Drescher日期:04/05/2018,浏览量:5976,Q&A: 0
One of the most exciting perspectives for studying bio-macromolecules comes from the emerging field of in-cell spectroscopy, which enables to determine the structure and dynamics of bio-macromolecules in the cell. In-cell electron paramagnetic resonance (EPR) spectroscopy in combination with micro-injection of bio-macromolecules into Xenopus ...
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癌症生物学

Primary Cultures from Human GH-secreting or Clinically Non-functioning Pituitary Adenomas 源自人类GH分泌的或临床上无功能的垂体腺瘤的原代培养物 作者:Roberto Würth, Alessandra Pattarozzi, Federica Barbieri and Tullio Florio日期:04/05/2018,浏览量:5359,Q&A: 0
Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological–based on somatostatin and dopamine agonists–approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary ...
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Centromere Chromosome Orientation Fluorescent in situ Hybridization (Cen-CO-FISH) Detects Sister Chromatid Exchange at the Centromere in Human Cells 着丝粒染色体定位荧光原位杂交法(Cen-CO-FISH)检测人细胞中着丝粒DNA重复的姐妹染色单体交换事件 作者:Simona Giunta日期:04/05/2018,浏览量:6648,Q&A: 0
Human centromeres are composed of large tandem arrays of repetitive alpha satellite DNA, which are often sites of aberrant rearrangement in cancers (Mitelman et al., 1997; Padilla-Nash et al., 2001). To date, annotation of the human centromere repetitive sequences remains incomplete, greatly hindering in-depth functional studies ...
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细胞生物学

Analysis of Mitochondrial Structure in the Body Wall Muscle of Caenorhabditis elegans 秀丽隐杆线虫体壁肌线粒体结构分析 作者:Shaarika Sarasija and Kenneth R. Norman日期:04/05/2018,浏览量:6882,Q&A: 0
Mitochondrial function is altered in various pathologies, highlighting the crucial role mitochondria plays in maintaining cellular homeostasis. Mitochondrial structure undergoes constant fission and fusion in response to changing cellular environment. Due to this, analyzing mitochondrial structure could provide insight into the physiological state ...
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发育生物学

A Microfluidic Device for Massively Parallel, Whole-lifespan Imaging of Single Fission Yeast Cells 一种用于单裂变酵母细胞大量平行的整个生命期成像的微流体装置 作者:Stephen K Jones Jr, Eric C Spivey, James R Rybarski and Ilya J Finkelstein日期:04/05/2018,浏览量:6592,Q&A: 0
Whole-lifespan single-cell analysis has greatly increased our understanding of fundamental cellular processes such as cellular aging. To observe individual cells across their entire lifespan, all progeny must be removed from the growth medium, typically via manual microdissection. However, manual microdissection is laborious, low-throughput, and ...
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免疫学

Imaging Cytokine Concentration Fields Using PlaneView Imaging Devices 使用PlaneView成像装置对细胞因子浓度视野进行成像 作者:Alon Oyler-Yaniv and Oleg Krichevsky日期:04/05/2018,浏览量:5315,Q&A: 0
We describe here a method to visualize concentration fields of cytokines around cytokine-secreting cells. The main challenge is that physiological cytokine concentrations can be very low, in the pico-molar range. Since it is currently impossible to measure such concentrations directly, we rely on cell’s response to the cytokines–the ...
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微生物学

Bacterial Competition Assay Based on Extracellular D-amino Acid Production 基于细胞外D-氨基酸生成的细菌竞争测定 作者:Laura Alvarez and Felipe Cava日期:04/05/2018,浏览量:6646,Q&A: 0
Bacteria live in polymicrobial communities under tough competition. To persist in a specific niche many species produce toxic extracellular effectors as a strategy to interfere with the growth of nearby microbes. One of such effectors are the non-canonical D-amino acids. Here we describe a method to test the effect of D-amino acid production in ...
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Purification of RNA Mango Tagged Native RNA-protein Complexes from Cellular Extracts Using TO1-Desthiobiotin Fluorophore Ligand 使用TO1-脱硫生物素荧光团配体从细胞提取物中纯化RNA Mango标记的天然RNA-蛋白质复合物 作者:Shanker Shyam Sundhar Panchapakesan, Sunny C. Y. Jeng and Peter J. Unrau日期:04/05/2018,浏览量:6177,Q&A: 2
A native purification strategy using RNA Mango for RNA based purification of RNA-protein complexes is described. The RNA Mango aptamer is first genetically engineered into the RNA of interest. RNA Mango containing complexes obtained from cleared cellular native extracts are then immobilized onto TO1-Desthiobiotin saturated streptavidin agarose ...
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神经科学

Context-driven Salt Seeking Test (Rats) 情景驱动的大鼠寻盐测试 作者:Stephen E. Chang and Kyle S. Smith日期:04/05/2018,浏览量:4601,Q&A: 0
Changes in reward seeking behavior often occur through incremental learning based on the difference between what is expected and what actually happens. Behavioral flexibility of this sort requires experience with rewards as better or worse than expected. However, there are some instances in which behavior can change through non-incremental ...
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Sacral Spinal Cord Transection and Isolated Sacral Cord Preparation to Study Chronic Spinal Cord Injury in Adult Mice 骶髓横断后制备骶髓分离物来研究成年小鼠慢性脊髓损伤 作者:Carmelo Bellardita, Maite Marcantoni, Peter Löw and Ole Kiehn日期:04/05/2018,浏览量:5698,Q&A: 0
Spinal cord injury (SCI) is characterized by multiple sensory/motor impairments that arise from different underlying neural mechanisms. Linking specific sensory/motor impairments to neural mechanism is limited by a lack of direct experimental access to these neural circuits. Here, we describe an experimental model which addresses this shortcoming. ...
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Assessing Prepulse Inhibition of Startle in Mice 评估小鼠惊吓的前脉冲抑制 作者:Christina Ioannidou, Giovanni Marsicano and Arnau Busquets-Garcia日期:04/05/2018,浏览量:9321,Q&A: 0
Animal models are an important tool for studying neuropsychiatric disorders. However, a major challenge for researchers working with laboratory rodents is trying to reproduce ‘core’ symptoms of complex human disorders such as schizophrenia. Despite this challenge, however, it is still conceivable to use animal models designed to reproduce some of ...
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Measuring Spatiotemporal Dynamics of Odor Gradient for Small Animals by Gas Chromatography 采用气相色谱法测定小动物对气味梯度的时空动力学 作者:Akiko Yamazoe-Umemoto, Yuishi Iwasaki and Koutarou D. Kimura日期:04/05/2018,浏览量:5560,Q&A: 0
Odor is the most fundamental chemical stimulus that delivers information regarding food, mating partners, enemies, and danger in the surrounding environment. Research on odor response in animals is widespread, although studies on experimental systems in which the gradient of odor concentration is quantitatively measured has been quite limited. ...
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Measurement of ROS in Caenorhabditis elegans Using a Reduced Form of Fluorescein 使用还原型荧光素测定秀丽隐杆线虫中的ROS 作者:Shaarika Sarasija and Kenneth R. Norman日期:04/05/2018,浏览量:4763,Q&A: 0
Oxidative stress is implicated in the pathogenesis of various neurodegenerative diseases, including Alzheimer’s disease. Oxidative stress is a result of a disruption of the equilibrium between antioxidants and oxidants, in favor of oxidants. Since mitochondria are major sites of production and reduction of reactive oxygen species (ROS), ...
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植物科学

Isolation of Nuclei in Tagged Cell Types (INTACT), RNA Extraction and Ribosomal RNA Degradation to Prepare Material for RNA-Seq 从标记的细胞类型中分离细胞核、提取RNA并去除核糖体RNA以用于RNA-Seq 分析 作者:Mauricio A. Reynoso, Germain C. Pauluzzi, Sean Cabanlit, Joel Velasco, Jérémie Bazin, Roger Deal, Siobhan Brady, Neelima Sinha, Julia Bailey-Serres and Kaisa Kajala日期:04/05/2018,浏览量:12219,Q&A: 0
Gene expression is dynamically regulated on many levels, including chromatin accessibility and transcription. In order to study these nuclear regulatory events, we describe our method to purify nuclei with Isolation of Nuclei in TAgged Cell Types (INTACT). As nuclear RNA is low in polyadenylated transcripts and conventional pulldown methods would ...
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In vitro Nitrate Reductase Activity Assay from Arabidopsis Crude Extracts 拟南芥粗提物中硝酸还原酶体外活性测定 作者:Joo Yong Kim and Hak Soo Seo日期:04/05/2018,浏览量:8122,Q&A: 0
Nitrate reductase (NR) reduces the major plant nitrogen source, NO3-, into NO2-. NR activity can be measured by its final product, nitrite through its absorbance under optimized condition. Here, we present a detailed protocol for measuring relative enzyme activity of NR from Arabidopsis crude ...
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Tracking Lipid Transfer by Fatty Acid Isotopolog Profiling from Host Plants to Arbuscular Mycorrhiza Fungi 脂肪酸同位素标记谱分析追踪脂质从寄主植物到丛枝菌根真菌的转运 作者:Andreas Keymer, Claudia Huber, Wolfgang Eisenreich and Caroline Gutjahr日期:04/05/2018,浏览量:6085,Q&A: 0
Lipid transfer from host plants to arbuscular mycorrhiza fungi was hypothesized for several years because sequenced arbuscular mycorrhiza fungal genomes lack genes encoding cytosolic fatty acid synthase (Wewer et al., 2014; Rich et al., 2017). It was finally shown by two independent experimental approaches (Jiang et al., ...
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Fluorescein Transport Assay to Assess Bulk Flow of Molecules Through the Hypocotyl in Arabidopsis thaliana 荧光素转运测定法评估通过拟南芥下胚轴的分子总体流动 作者:Salva Duran-Nebreda and George W. Bassel日期:04/05/2018,浏览量:5235,Q&A: 0
The bulk transport of molecules through plant tissues underpins growth and development. The stem acts as a conduit between the upper and low domains of the plant, facilitating transport of solutes and water from the roots to the shoot system, and sugar plus other elaborated metabolites towards the non-photosynthetic organs. In order to perform ...
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RNA Purification from the Unicellular Green Alga, Chromochloris zofingiensis 单细胞绿藻佐夫色绿藻中的RNA纯化 作者:Sean D. Gallaher and Melissa S. Roth日期:04/05/2018,浏览量:5915,Q&A: 0
Chromochloris zofingiensis is a unicellular green alga that is an emerging model species for studies in fields such as biofuel production, ketocarotenoid biosynthesis and metabolism. The recent availability of a high-quality genome assembly facilitates systems-level analysis, such as RNA-Seq. However, cells of this alga have a tough cell ...
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Evaluation of Root pH Change Through Gel Containing pH-sensitive Indicator Bromocresol Purple 通过含有pH敏感指示剂溴甲酚紫的凝胶评估根部pH值的变化 作者:Aparecida L. Silva, Keini Dressano, Paulo H. O. Ceciliato, Juan Carlos Guerrero-Abad and Daniel S. Moura日期:04/05/2018,浏览量:7020,Q&A: 1
The Rapid Alkalinization Factor (RALF) is a plant hormone peptide that inhibits proton transport causing alkalinization of the extracellular media. To detect the alkalinization response elicited by RALF peptides in root cells, Arabidopsis seedlings are carefully transferred to a gel containing the pH-sensitive indicator bromocresol ...
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干细胞

Generating Loss-of-function iPSC Lines with Combined CRISPR Indel Formation and Reprogramming from Human Fibroblasts 人成纤维细胞通过CRISPR 插入缺失的形成联合重编程生成功能丧失iPSC细胞系 作者:Andrew M. Tidball, Preethi Swaminathan, Louis T. Dang and Jack M. Parent日期:04/05/2018,浏览量:9279,Q&A: 1
For both disease and basic science research, loss-of-function (LOF) mutations are vitally important. Herein, we provide a simple stream-lined protocol for generating LOF iPSC lines that circumvents the technical challenges of traditional gene-editing and cloning of established iPSC lines by combining the introduction of the CRISPR vector ...
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