• 卷册 11, 2021
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Image Credit: BioProtoc.4105

生物化学

Efficient and Rapid Analysis of Polysomes and Ribosomal Subunits in Cells and Tissues Using Ribo Mega-SEC 利用Ribo Mega-SEC高效快速分析细胞和组织中的多聚体和核糖体亚基 作者:Harunori Yoshikawa, Ramasubramanian Sundaramoorthy, Daniel Mariyappa, Hao Jiang and Angus I. Lamond日期:08/05/2021,浏览量:1610,Q&A: 0

Polysome profile analysis is a popular method for separating polysomes and ribosomal subunits and is typically achieved using a sucrose density gradient (SDG). This has remained the gold standard method since ribosomes were first discovered; however, this method is time-consuming and requires multiple steps from making the gradient and long

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Purification of Mitochondrial Ribosomes with the Translocase Oxa1L from HEK Cells 用转位酶Oxa1L纯化HEK细胞线粒体核糖体 作者:Hanting Yang and Nirupa Desai日期:08/05/2021,浏览量:1323,Q&A: 0

Mitochondrial ribosomes (mitoribosomes) perform protein synthesis inside mitochondria, the organelles responsible for energy conversion and adenosine triphosphate (ATP) production in eukaryotic cells. To investigate their functions and structures, large-scale purification of intact mitoribosomes from mitochondria-rich animal tissues or HEK cells

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Purification of Recombinant Wild Type and Mutant Ryanodine Receptors Expressed in HEK293 Cells HEK293细胞表达的重组野生型和突变型Ryanodine受体的纯化 作者:Yifan Hu, Kavita A. Iyer, Ashok R. Nayak, Nagomi Kurebayashi, Takashi Murayama and Montserrat Samsó日期:08/05/2021,浏览量:1054,Q&A: 0

High quantities of purified ryanodine receptor (RyR), a large (2.26 MDa) intracellular homotetrameric membrane protein, can be obtained from heterologous expression in HEK293 cells and used for structure determination by cryo-EM. The advantage of using recombinant protein is that the variability due to post-translational modifications can be

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生物物理学

Purification and Cryo-electron Microscopy Analysis of Plant Mitochondrial Ribosomes 植物线粒体核糖体的纯化和低温电镜分析 作者:Florent Waltz, Philippe Giegé and Yaser Hashem日期:08/05/2021,浏览量:1038,Q&A: 0

Plants make up by far the largest part of biomass on Earth. They are the primary source of food and the basis of most drugs used for medicinal purposes. Similarly to all eukaryotes, plant cells also use mitochondria for energy production. Among mitochondrial gene expression processes, translation is the least understood; although, recent advances

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细胞生物学

CRISPR-mediated Labeling of Cells in Chick Embryos Based on Selectively Expressed Genes 基于选择性表达基因的CRISPR介导的鸡胚胎细胞标记 作者:Masahito Yamagata and Joshua R. Sanes日期:08/05/2021,浏览量:1577,Q&A: 0

The abilities to mark and manipulate specific cell types are essential for an increasing number of functional, structural, molecular, and developmental analyses in model organisms. In a few species, this can be accomplished by germline transgenesis; in other species, other methods are needed to selectively label somatic cells based on the genes

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Cell-attached and Whole-cell Patch-clamp Recordings of Dopamine Neurons in the Substantia Nigra Pars Compacta of Mouse Brain Slices 小鼠脑片黑质致密部多巴胺神经元的细胞贴附和全细胞膜片钳记录 作者:Stefano Cattaneo, Maria Regoni, Jenny Sassone and Stefano Taverna日期:08/05/2021,浏览量:1072,Q&A: 0

The Substantia Nigra pars compacta (SNc) is a midbrain dopaminergic nucleus that plays a key role in modulating motor and cognitive functions. It is crucially involved in several disorders, particularly Parkinson’s disease, which is characterized by a progressive loss of SNc dopaminergic cells. Electrophysiological studies on SNc neurons are

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发育生物学

Isolation and in vitro Culture of Mouse Oocytes 小鼠卵母细胞的分离与体外培养 作者:Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer日期:08/05/2021,浏览量:1258,Q&A: 0

Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success in fertility clinics. In vitro culture systems enable researchers to analyse important molecular and

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A Genetically Engineered Mouse Model of Venous Anomaly and Retinal Angioma-like Vascular Malformation 静脉畸形和视网膜血管瘤样血管畸形的基因工程小鼠模型 作者:Xudong Cao, Beibei Xu, Xiao Li, Taotao Li and Yulong He日期:08/05/2021,浏览量:1070,Q&A: 0

Characterization of key regulators in vein development will advance our understanding of mechanisms underlying venous anomalies and provide therapeutic targets for the treatment of vascular malformations. Here, we provide a detailed protocol for the generation of genetically engineered mouse models targeting the Tek gene for the analysis of vein

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医学

A Swimming-based Assay to Determine the Exercise Capacity of Adult Zebrafish Cardiomyopathy Models 基于游泳的测定成年斑马鱼心肌病模型运动能力的方法 作者:Xiao Ma and Xiaolei Xu日期:08/05/2021,浏览量:844,Q&A: 0

Exercise capacity, measured by treadmill in humans and other mammals, is an important diagnostic and prognostic index for patients with cardiomyopathy and heart failure. The adult zebrafish is increasingly used as a vertebrate model to study human cardiomyopathy due to its conserved cardiovascular physiology, convenience for genetic manipulation,

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微生物学

Ion Transport Activity Assay for Microbial Rhodopsin Expressed in Escherichia coli Cells 大肠杆菌表达的微生物紫红质离子转运活性测定 作者:Masae Konno, Keiichi Inoue and Hideki Kandori日期:08/05/2021,浏览量:878,Q&A: 0

Microbial rhodopsins have diverse functions, including roles as light-driven ion pumps, light-gated ion channels, photosensors, and light-regulated enzymes. As the number of rhodopsin-like genes identified has increased in recent years, so has the requirement for rapid identification of their functions. The patch-clamp method is often used to

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分子生物学

Isolation of Nuclei from Mouse Dorsal Root Ganglia for Single-nucleus Genomics 小鼠背根神经节细胞核的分离及其单核基因组学研究 作者:Lite Yang, Ivan Tochitsky, Clifford J. Woolf and William Renthal日期:08/05/2021,浏览量:980,Q&A: 0

Primary somatosensory neurons, whose cell bodies reside in the dorsal root ganglion (DRG) and trigeminal ganglion, are specialized to transmit sensory information from the periphery to the central nervous system. Our molecular understanding of peripheral sensory neurons has been limited by both their heterogeneity and low abundance compared with

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Fe-NTA Microcolumn Purification of Phosphopeptides from Immunoprecipitation (IP) Eluates for Mass Spectrometry Analysis Fe-NTA微柱纯化免疫沉淀(IP)洗脱物中磷酸肽的质谱分析 作者:Ethan J. Sanford and Marcus B. Smolka日期:08/05/2021,浏览量:798,Q&A: 0

Protein phosphorylation is a nearly universal signaling mechanism. To date, a number of proteomics tools have been developed to analyze phosphorylation. Phosphoproteome-wide analyses using whole cell extracts suffer from incomplete coverage, often missing phosphorylation events from low-abundance proteins. In order to increase coverage of

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神经科学

Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata 小鼠和人胰腺的光学清除和三维分析优化 作者:Alexandra Alvarsson, Maria Jimenez-Gonzalez, Rosemary Li, Carolina Rosselot, Nikolaos Tzavaras, Zhuhao Wu and Sarah A. Stanley日期:08/05/2021,浏览量:1591,Q&A: 0

The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have allowed detailed reconstructions of pancreatic innervation. Optical clearing is used to enhance tissue

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A Simple Spatial-independent Associative and Reversal Learning Task in Mice 小鼠的一个简单的空间独立的联想和逆转学习任务 作者:Fulvio Magara, Benjamin Boury-Jamot and Hanna Hörnberg日期:08/05/2021,浏览量:767,Q&A: 0

The ability to adapt one's behavior in response to changing circumstances, or cognitive flexibility, is often altered in neuropsychiatric and neurodevelopmental conditions. In rodents, cognitive flexibility is frequently assessed using associative learning paradigms with a reversal component. The majority of existing protocols rely on

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植物科学

A Fast and Easy Method to Study Ralstonia solanacearum Virulence upon Transient Gene Expression or Gene Silencing in Nicotiana benthamiana Leaves 利用本生烟叶片基因瞬时表达或沉默研究青枯菌毒力的快速简便方法 作者:Wenjia Yu and Alberto P. Macho日期:08/05/2021,浏览量:1161,Q&A: 0

Ralstonia solanacearum is a devastating soil-borne bacterial pathogen that causes disease in multiple host plants worldwide. Typical assays to measure virulence of R. solanacearum in laboratory conditions rely on soil-drenching inoculation followed by observation and scoring of disease symptoms. Here, we describe a novel inoculation protocol to

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干细胞

Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures 小鼠骨膜细胞培养、体外分化及胫骨骨折的体内移植 作者:Simon Perrin, Anais Julien, Oriane Duchamp de Lageneste, Rana Abou-Khalil and Céline Colnot日期:08/05/2021,浏览量:925,Q&A: 0

The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or enzymatic digestion. Here, we describe an explant culture method to isolate periosteum-derived

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