• 卷册 11, 2021
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Image Credit: BioProtoc.4209

生物化学

Thermal Proteome Profiling to Identify Protein-ligand Interactions in the Apicomplexan Parasite Toxoplasma gondii 弓形虫热蛋白质组分析以识别顶复体寄生虫中的蛋白质-配体相互作用 作者:Alice L. Herneisen and Sebastian Lourido日期:11/05/2021,浏览量:1265,Q&A: 0

Toxoplasma gondii is a single-celled eukaryotic parasite that chronically infects a quarter of the global population. In recent years, phenotypic screens have identified compounds that block parasite replication. Unraveling the pathways and molecular mechanisms perturbed by such compounds requires target deconvolution. In parasites, such

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生物工程

Development of a 3D Bioprinted Scaffold with Spatio-temporally Defined Patterns of BMP-2 and VEGF for the Regeneration of Large Bone Defects 开发具有 BMP-2 和 VEGF 时空定义模式的 3D 生物打印支架,用于大骨缺损的再生 作者:Fiona E. Freeman, Pierluca Pitacco, Lieke H. A. van Dommelen, Jessica Nulty, David C. Browe, Jung-Youn Shin, Eben Alsberg and Daniel J. Kelly日期:11/05/2021,浏览量:931,Q&A: 0

The local delivery of growth factors such as BMP-2 is a well-established strategy for the repair of bone defects. The limitations of such approaches clinically are well documented and can be linked to the need for supraphysiological doses and poor spatio-temporal control of growth factor release in vivo. Using bioprinting techniques, it is

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癌症生物学

Measuring Real-time DNA/RNA Nuclease Activity through Fluorescence 通过荧光测量实时 DNA/RNA 核酸酶活性 作者:Paulina Wyrzykowska, Sally Rogers and Richard Chahwan日期:11/05/2021,浏览量:1008,Q&A: 0

DNA and RNA nucleases are wide-ranging enzymes, taking part in broad cellular processes from DNA repair to immune response control. Growing interest in the mechanisms and activities of newly discovered nucleases inspired us to share the detailed protocol of our nuclease assay (Sheppard et al., 2019). This easy and inexpensive method can provide

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发育生物学

Intact in situ Preparation of the Drosophila melanogaster Lymph Gland for a Comprehensive Analysis of Larval Hematopoiesis 黑腹果蝇淋巴腺的完整原位制备用于综合幼虫造血分析 作者:Diana Rodrigues, K. VijayRaghavan, Lucas Waltzer and Maneesha S. Inamdar日期:11/05/2021,浏览量:1304,Q&A: 0

Blood cells have a limited lifespan and are replenished by a small number of hematopoietic stem and progenitor cells (HSPCs). Adult vertebrate hematopoiesis occurs in the bone marrow, liver, and spleen, rendering a comprehensive analysis of the entire HSPC pool nearly impossible. The Drosophila blood system is well studied and has developmental,

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Targeting the Expression of Long Noncoding RNAs in Murine Satellite Cells from Single Myofibers 靶向小鼠卫星细胞中长链非编码 RNA 的表达 作者:Martina Macino, Beatrice Biferali, Andrea Cipriano, Monica Ballarino and Chiara Mozzetta日期:11/05/2021,浏览量:853,Q&A: 0

LncRNAs have been recently implicated in the epigenetic control of muscle differentiation and their functional characterization has traditionally relied upon in vitro models of myogenic differentiation. However, the use of experimental paradigms to specifically target lncRNAs expression in muscle stem cells (MuSCs), also known as satellite cells,

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APEX-mediated Proximity Labeling of Proteins in Cells Targeted by Extracellular Vesicles APEX 介导的细胞外囊泡靶向细胞中蛋白质的邻近标记 作者:Lu Song, Jun Chen, Angela Sun and Randy Schekman日期:11/05/2021,浏览量:962,Q&A: 0

Extracellular vesicles (EVs) are thought to mediate intercellular communication through the delivery of cargo proteins and RNA to target cells. The uptake of EVs is often followed visually using lipophilic-dyes or fluorescently-tagged proteins to label membrane constituents that are then internalized into recipient cells (Christianson et al.,

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免疫学

Lentivirus-mediated Conditional Gene Expression 慢病毒介导的条件基因表达 作者:Leah Rommereim, Ajay Suresh Akhade, Ronald N. Germain, Iain D. C. Fraser and Naeha Subramanian日期:11/05/2021,浏览量:908,Q&A: 0

The ability to identify the role of a particular gene within a system is dependent on control of the expression of that gene. In this protocol, we describe a method for stable, conditional expression of Nod-Like receptors (NLRs) in THP-1 cells using a lentiviral expression system. This system combines all the necessary components for

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Suppression of Human Dendritic Cells by Regulatory T Cells 调节性 T 细胞抑制人树突状细胞 作者:Qing Huang, Avery J. Lam, Dominic A. Boardman, Nicholas A. J. Dawson and Megan K. Levings日期:11/05/2021,浏览量:670,Q&A: 0

Regulatory T cells (Tregs) suppress immune responses via a variety of mechanisms and can be used as a cellular therapy to induce tolerance. The function of Tregs is commonly assessed in vitro using assays that measure suppression of effector T cell proliferation and/or cytokine production. However, Tregs can also suppress the function of antigen

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Production of Recombinant Replication-defective Lentiviruses Bearing the SARS-CoV or SARS-CoV-2 Attachment Spike Glycoprotein and Their Application in Receptor Tropism and Neutralisation Assays 携带 SARS-CoV 或 SARS-CoV-2 附着尖峰糖蛋白的重组复制缺陷慢病毒的生产及其在受体向性和中和试验中的应用 作者:Nazia Thakur, Giulia Gallo, Ahmed M. E. Elreafey and Dalan Bailey日期:11/05/2021,浏览量:632,Q&A: 0

For enveloped viruses, such as SARS-CoV-2, transmission relies on the binding of viral glycoproteins to cellular receptors. Conventionally, this process is recapitulated in the lab by infection of cells with isolated live virus. However, such studies can be restricted due to the availability of high quantities of replication-competent virus,

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微生物学

An Assay to Determine NAD(P)H: Quinone Oxidoreductase Activity in Cell Extracts from Candida glabrata 测定 NAD(P)H 的分析:光滑念珠菌细胞提取物中的醌氧化还原酶活性 作者:Anamika Battu, Rajaram Purushotham and Rupinder Kaur日期:11/05/2021,浏览量:469,Q&A: 0

Flavodoxin-like proteins (Fld-LPs) are an important constituent of the oxidative stress defense system in several organisms and highly conserved from bacteria to humans. These proteins possess NAD(P)H:quinone oxidoreductase activity and convert quinones to hydroquinones through two-electron reduction, using NAD(P)H and quinone as electron donor

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Quantification of Duloxetine in the Bacterial Culture and Medium to Study Drug-gut Microbiome Interactions 在细菌培养和培养基中定量度洛西汀以研究药物-肠道微生物组的相互作用 作者:Prasad B. Phapale, Sonja Blasche, Kiran R. Patil and Theodore Alexandrov日期:11/05/2021,浏览量:465,Q&A: 0

Expanding our understanding of drug-gut bacteria interactions requires high-throughput drug measurements in complex bacterial cultures. Quantification of drugs in the cultures, media, and cell pellets is prone to strong matrix effects. We have developed a liquid chromatography–high resolution mass spectrometry (LC–HRMS) method for

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Determination of Fungal Tolerance Index to Heavy Metals and Heavy Metal Resistance Tests 真菌对重金属的耐受指数的测定和重金属抗性试验 作者:Rosa Paulina Calvillo-Medina日期:11/05/2021,浏览量:453,Q&A: 0

Fungal metallo-tolerance has been described in different species and plays an important role in bioremediation of contaminated environments. Metallo-tolerance is mainly documented by microdilution assays and agar well diffusion methods using equipment that can be expensive. The tolerance index can be calculated to determine the efficiency of a

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分子生物学

Enrichment of Cytoplasmic RNA Granules from Arabidopsis Seedlings 从拟南芥幼苗中富集细胞质 RNA 颗粒 作者:Zhen Lei, Eun Yu Kim and Jungnam Cho日期:11/05/2021,浏览量:522,Q&A: 0

RNA granules (RGs) are membraneless intracellular compartments that play important roles in the post-transcriptional control of gene expression. Stress granules (SGs) are a type of RGs that form under environmental challenges and/or internal cellular stresses. Stress treatments lead to strong mRNAs translational inhibition and storage in SGs until

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Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes 严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) 慢病毒假型的生产、滴定、中和、储存和冻干 作者:Cecilia Di Genova, Alex Sampson, Simon Scott, Diego Cantoni, Martin Mayora-Neto, Emma Bentley, Giada Mattiuzzo, Edward Wright, Mariliza Derveni, Bethany Auld, Bill T. Ferrara, Dale Harrison, Mohamed Said, Arwa Selim, Elinor Thompson, Craig Thompson, George Carnell and Nigel Temperton日期:11/05/2021,浏览量:702,Q&A: 0

This protocol details a rapid and reliable method for the production and titration of high-titre viral pseudotype particles with the SARS-CoV-2 spike protein (and D614G or other variants of concern, VOC) on a lentiviral vector core, and use for neutralisation assays in target cells expressing angiotensin-converting enzyme 2 (ACE2) and

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A Nucleocapsid-based Transcomplementation Cell Culture System of SARS-CoV-2 to Recapitulate the Complete Viral Life Cycle 基于核衣壳的 SARS-CoV-2 转互补细胞培养系统,重现完整的病毒生命周期 作者:Yanying Yu, Xiaohui Ju and Qiang Ding日期:11/05/2021,浏览量:411,Q&A: 0

The ongoing COVID-19 pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). As this virus is classified as a biosafety level-3 (BSL-3) agent, the development of countermeasures and basic research methods is logistically difficult. Recently, using reverse genetics, we developed a BSL-2 cell culture system for production

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神经科学

Whole-mount Staining of Mouse Diaphragm Neuromuscular Junctions 小鼠膈肌神经肌肉接头的整体染色 作者:Rula Sha, Zhenzhen Wang, Xue You, Yuguo Liu, Zhiqin Xie and Ying Feng日期:11/05/2021,浏览量:410,Q&A: 0

The neuromuscular junction (NMJ) is a specialized synapse that connects the terminal end of a motor neuron and a skeletal muscle fiber. Defects in NMJ cause abnormalities of neuromuscular transmission, leading to NMJ disorders. The mammalian diaphragm muscle is essential for respiration and has been widely used to study NMJ formation. Here, we

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植物科学

Preference Test of Plutella xylostella Larvae upon DMNT Treatment 小菜蛾幼虫对DMNT处理的偏好试验 作者:Chen Chen, Zhen Tao, Chuanhong Wang, Tengyue Wang, Hongyi Chen, Taoshan Jiang and Peijin Li日期:11/05/2021,浏览量:389,Q&A: 0

We describe a method to test the preference of insects in response to (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). We use a device that includes a horizontal glass tube, two grooves (with activated carbon), air flow, rubber stoppers/tubes, transparent glass containers (optional), and a holder for the glass tube (optional). Equal amounts of activated

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干细胞

High-throughput 3D Spheroid Formation and Effective Cardiomyocyte Differentiation from Human iPS Cells Using the Microfabric Vessels EZSPHERETM 利用微细血管EZSPHERETM从人类iPS细胞中进行高通量三维球状细胞形成和有效的心肌细胞分化 作者:Tatsuaki Miwa, Alimjan Idiris and Hiromichi Kumagai日期:11/05/2021,浏览量:514,Q&A: 0

High-throughput 3D spheroid formation from human induced pluripotent stem cells (hiPSCs) can be easily performed using the unique microfabric vessels EZSPHERE, resulting in effective and large scale generation of differentiated cells such as cardiomyocytes or neurons. Such hiPSC-derived cardiomyocytes (hiPSC-CMs) or neurons are very useful in the

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系统生物学

Histone modification ChIP-seq on Arabidopsis thaliana Plantlets 拟南芥植株上的组蛋白修饰ChIP-seq 作者:Juan S. Ramirez-Prado, David Latrasse and Moussa Benhamed日期:11/05/2021,浏览量:588,Q&A: 1

Characterizing the molecular mechanisms regulating gene expression is crucial for understanding the regulatory processes underlying physiological responses to environmental and developmental signals in eukaryotes. The covalent modification of histones contributes to the compaction levels of chromatin, as well as the recruitment of the

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