• 卷册 11, 2021
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Image Credit: BioProtoc.4150

往期刊物(2021)

卷册: 11 期号: 17

生物化学

Kinetic Analysis of a Protein-protein Complex to Determine its Dissociation Constant (KD) and the Effective Concentration (EC50) of an Interplaying Effector Molecule Using Bio-layer Interferometry 蛋白质-蛋白质复合物的动力学分析,以确定其电离常数 (KD) 和使用生物层干涉法相互作用的效应分子的有效浓度 (EC50) 作者:Tim Orthwein, Luciano F. Huergo, Karl Forchhammer and Khaled A. Selim日期:09/05/2021,浏览量:1167,Q&A: 0

Biolayer interferometry (BLI) is an emerging analytical tool that allows the study of protein complexes in real time to determine protein complex kinetic parameters. This article describes a protocol to determine the KD of a protein complex using a 6×His tagged fusion protein as bait immobilized on the NTA sensor chip of the

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生物物理学

Using Atomic Force Microscopy to Study the Real Time Dynamics of DNA Unwinding by Mitochondrial Twinkle Helicase 使用原子力显微镜研究线粒体闪烁解旋酶对DNA解旋的实时动力学 作者:Parminder Kaur, Hai Pan, Matthew J. Longley, William C. Copeland and Hong Wang日期:09/05/2021,浏览量:906,Q&A: 0

Understanding the structure and dynamics of DNA-protein interactions during DNA replication is crucial for elucidating the origins of disorders arising from its dysfunction. In this study, we employed Atomic Force Microscopy as a single-molecule imaging tool to examine the mitochondrial DNA helicase Twinkle and its interactions with DNA. We used

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细胞生物学

Cytoduction and Plasmiduction in Yeast 酵母细胞诱导中的细胞诱导和质粒诱导 作者:Jane E. Dorweiler and Anita L. Manogaran日期:09/05/2021,浏览量:846,Q&A: 0

Cytoduction, and a related technique referred to as plasmiduction, have facilitated substantial advancements in the field of yeast prion biology by providing a streamlined method of transferring prions from one yeast strain to another. Prions are cytoplasmic elements consisting of aggregated misfolded proteins, and as such, they exhibit

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发育生物学

Isolation of Myofibres and Culture of Muscle Stem Cells from Adult Zebrafish 成年斑马鱼肌纤维的分离和肌肉干细胞的培养 作者:Massimo Ganassi, Peter S. Zammit and Simon M. Hughes日期:09/05/2021,浏览量:1383,Q&A: 0

Skeletal muscles generate force throughout life and require maintenance and repair to ensure efficiency. The population of resident muscle stem cells (MuSCs), termed satellite cells, dwells beneath the basal lamina of adult myofibres and contributes to both muscle growth and regeneration. Upon exposure to activating signals, MuSCs proliferate to

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免疫学

Plasmodium cynomolgi Berok Growth Inhibition Assay by Thiol-reactive Probe Based Flow Cytometric Measurement 硫醇反应探针流式细胞术检测食蟹猴疟原虫生长抑制 作者:Jessica Jie Ying Ong, Bruce Russell and Jin-Hee Han日期:09/05/2021,浏览量:758,Q&A: 0

The relapsing malaria species, Plasmodium vivax, is the most widely distributed and difficult-to-treat cause of human malaria. The merozoites of P. vivax preferentially invade ephemeral human CD71+ reticulocytes (nascent reticulocytes), thereby limiting the development of a robust continuous culture in vitro. Fortunately, P. vivax’s sister

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Isolation and Quantification of Mouse γδT-cells in vitro and in vivo 体外和体内小鼠γδT细胞的分离和定量 作者:Isha Rana, Krithika Badarinath, Ravindra K. Zirmire and Colin Jamora日期:09/05/2021,浏览量:974,Q&A: 0

The skin plays an important role in protecting the body from pathogens and chemicals in the external environment. Upon injury, a healing program is rapidly initiated and involves extensive intercellular communication to restore tissue homeostasis. The deregulation of this crosstalk can lead to abnormal healing processes and is the foundation of

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Microscopic Detection of ASC Inflammasomes in Bone Marrow Derived Macrophages Post Stimulation 刺激后骨髓源性巨噬细胞中ASC炎症小体的显微检测 作者:Longjun Li, Rudi Mao and Xiaopeng Qi日期:09/05/2021,浏览量:840,Q&A: 0

An inflammasome is an intracellular multiprotein complex that plays important roles in host defense and inflammatory responses. Inflammasomes are typically composed of the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), cytoplasmic sensor protein, and the effector protein pro-caspase-1. ASC assembly into a protein

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微生物学

Isolation and Characterization of Membrane Vesicles from Lactobacillus Species 乳酸杆菌属膜囊泡中的分离和特性研究 作者:Julie C. Caruana, Scott N. Dean and Scott A. Walper日期:09/05/2021,浏览量:937,Q&A: 0

Throughout their life cycle, bacteria shed portions of their outermost membrane comprised of proteins, lipids, and a diversity of other biomolecules. These biological nanoparticles have been shown to have a range of highly diverse biological activities, including pathogenesis, community regulation, and cellular defense (among others). In recent

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分子生物学

Direct-TRI: High-throughput RNA-extracting Method for all Stages of Zebrafish Development Direct-TRI: 斑马鱼发育各阶段高通量RNA提取方法 作者:Kota Ujibe, Kanako Nishimura, Makoto Kashima and Hiromi Hirata日期:09/05/2021,浏览量:614,Q&A: 0

Recent popularization of next-generation sequencing enables conducting easy transcriptome analysis. Nevertheless, substantial RNA isolation work prior to RNA sequencing, as well as the high cost involved, still makes the routine use of large-scale transcriptome analysis difficult. For example, conventional phenol-chloroform RNA extraction cannot

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In vitro Cleavage and Electrophoretic Mobility Shift Assays for Very Fast CRISPR 快速CRISPR的体外裂解和电泳迁移率位移测定 作者:Roger S. Zou, Yang Liu and Taekjip Ha日期:09/05/2021,浏览量:867,Q&A: 0

CRISPR-Cas9 has transformed biomedical research and medicine through convenient and targeted manipulation of DNA. Time- and spatially-resolved control over Cas9 activity through the recently developed very fast CRISPR (vfCRISPR) system have facilitated comprehensive studies of DNA damage and repair. Understanding the fundamental principles of Cas9

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Preparation and Characterization of Internally Modified DNA Templates for Chemical Transcription Roadblocking 用于化学转录障碍的内部修饰 DNA 模板的制备和特征分析 作者:Eric J. Strobel日期:09/05/2021,浏览量:484,Q&A: 0

Site-specific transcription arrest is the basis of emerging technologies that assess nascent RNA structure and function. Cotranscriptionally folded RNA can be displayed from an arrested RNA polymerase (RNAP) for biochemical manipulations by halting transcription elongation at a defined DNA template position. Most transcription

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神经科学

Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay 通过邻近连接测定法测量 LRRK2 激酶活性 作者:Matthew T. Keeney, Eric K. Hoffman, J. Timothy Greenamyre and Roberto Di Maio日期:09/05/2021,浏览量:669,Q&A: 0

Missense mutations in leucine rich-repeat kinase 2 (LRRK2) cause forms of familial Parkinson’s disease and have been linked to ‘idiopathic’ Parkinson’s disease. Assessment of LRRK2 kinase activity has been very challenging due to its size, complex structure, and relatively low abundance. A standard in the field to assess

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植物科学

U2.3 Precursor Small Nuclear RNA in vitro Processing Assay U2.3前体小核 RNA体外加工试验 作者:Chan Lin, Yujie Feng, Xueyan Peng, Jiaming Wu, Weili Wang and Yunfeng Liu日期:09/05/2021,浏览量:390,Q&A: 0

Small nuclear RNAs (snRNAs) are vital for eukaryotic cell activities and play important roles in pre-mRNA splicing. The molecular mechanism underlying the transcription of snRNA, regulated via upstream/downstream cis-elements and relevant trans-elements, has been investigated in detail using cell-free extracts. However, the processing of precursor

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Solubilization Method for Isolation of Photosynthetic Mega- and Super-complexes from Conifer Thylakoids 从针叶树类囊体中分离光合巨复合体和超复合体的增溶方法 作者:Pushan Bag, Wolfgang P. Schröder, Stefan Jansson and Domenica Farci日期:09/05/2021,浏览量:639,Q&A: 0

Photosynthesis is the main process by which sunlight is harvested and converted into chemical energy and has been a focal point of fundamental research in plant biology for decades. In higher plants, the process takes place in the thylakoid membranes where the two photosystems (PSI and PSII) are located. In the past few decades, the evolution of

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干细胞

In situ Hybridization of miRNAs in Human Embryonic Kidney and Human Pluripotent Stem Cell-derived Kidney Organoids 在人胚肾和人多能干细胞衍生的肾类器官中微小RNA的原位杂交 作者:Filipa M. Lopes, Susan J. Kimber and Ioannis Bantounas日期:09/05/2021,浏览量:539,Q&A: 0

MicroRNAs are small RNAs that negatively regulate gene expression and play an important role in fine-tuning molecular pathways during development. There is increasing interest in studying their function in the kidney, but the majority of studies to date use kidney cell lines and assess the total amounts of miRNAs of interest either by qPCR or by

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