• 卷册 11, 2021
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Image Credit: BioProtoc.4100

现期刊物(2021)

卷册: 11, 期号: 14

生物化学

A Gel-Based Assay for Probing Protein Translocation on dsDNA 一种基于凝胶的检测dsDNA蛋白易位的方法 作者:Christiane Brugger and Alexandra M. Deaconescu日期:07/20/2021,浏览量:87,Q&A: 0

Protein translocation on DNA represents the key biochemical activity of ssDNA translocases (aka helicases) and dsDNA translocases such as chromatin remodelers. Translocation depends on DNA binding but is a distinct process as it typically involves multiple DNA binding states, which are usually dependent on nucleotide binding/hydrolysis and are

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Protocol for Spontaneous and Chaperonin-assisted in vitro Refolding of a Slow-folding Mutant of GFP, sGFP 一个GFP慢折叠突变体的自发和伴侣蛋白辅助的体外重折叠方案 作者:Anwar Sadat, Satyam Tiwari and Koyeli Mapa日期:07/20/2021,浏览量:83,Q&A: 0

Understanding the folding pathway of any protein is of utmost importance for deciphering the folding problems under adverse conditions. We can obtain important information about the folding pathway by monitoring the folding of any protein from its unfolded state. It is usually very difficult to monitor the folding process in real time as the

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生物物理学

A Multi-color Bicistronic Biosensor to Compare the Translation Dynamics of Different Open Reading Frames at Single-molecule Resolution in Live Cells 一种多色双顺反子生物传感器比较活细胞中不同开放阅读框在单分子分辨率下的翻译动力学 作者:Amanda L. Koch, Tatsuya Morisaki and Timothy J. Stasevich日期:07/20/2021,浏览量:73,Q&A: 0

Here, we describe how to image and quantitate the translation dynamics of a bicistronic biosensor that we recently created to fairly compare cap-dependent and IRES-mediated translation at single-molecule resolution in live human cells. This technique employs a pair of complementary intrabodies loaded into living cells that co-translationally bind

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Modeling Perturbations in Protein Filaments at the Micro and Meso Scale Using NAMD and PTools/Heligeom 用NAMD和PTools/Heligeom模拟蛋白质丝的微尺度和中尺度扰动 作者:Benjamin Boyer, Benoist Laurent, Charles H. Robert and Chantal Prévost日期:07/20/2021,浏览量:179,Q&A: 0

Protein filaments are dynamic entities that respond to external stimuli by slightly or substantially modifying the internal binding geometries between successive protomers. This results in overall changes in the filament architecture, which are difficult to model due to the helical character of the system. Here, we describe how distortions in RecA

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癌症生物学

Modeling the Nonlinear Dynamics of Intracellular Signaling Networks 细胞内信号网络的非线性动力学建模 作者:Oleksii S. Rukhlenko and Boris N. Kholodenko日期:07/20/2021,浏览量:113,Q&A: 0

This protocol illustrates a pipeline for modeling the nonlinear behavior of intracellular signaling pathways. At fixed spatial points, nonlinear signaling dynamics are described by ordinary differential equations (ODEs). At constant parameters, these ODEs may have multiple attractors, such as multiple steady states or limit cycles. Standard

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细胞生物学

Preparation of Human Chondrocytes for Profiling Using Cytometry by Time-of-flight (cyTOF) 用飞行时间(cyTOF flight)流式细胞术制备人软骨细胞用于分析研究 作者:Fiorella Carla Grandi and Nidhi Bhutani日期:07/20/2021,浏览量:84,Q&A: 0

Single-cell technologies have allowed high-resolution profiling of tissues and thus a deeper understanding of tissue homeostasis and disease heterogeneity. Understanding this heterogeneity can be especially important for tailoring treatments in a patient-specific manner. Here, we detail methods for preparing human cartilage tissue for profiling

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Analysis of the Effects of Hexokinase 2 Detachment From Mitochondria-Associated Membranes with the Highly Selective Peptide HK2pep 用高选择性肽HK2pep分析己糖激酶2脱离线粒体相关膜的作用 作者:Francesco Ciscato, Federica Chiara, Riccardo Filadi and Andrea Rasola日期:07/20/2021,浏览量:97,Q&A: 0

The crucial role of hexokinase 2 (HK2) in the metabolic rewiring of tumors is now well established, which makes it a suitable target for the design of novel therapies. However, hexokinase activity is central to glucose utilization in all tissues; thus, enzymatic inhibition of HK2 can induce severe adverse effects. In an effort to find a selective

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发育生物学

GeneWeld: Efficient Targeted Integration Directed by Short Homology in Zebrafish GeneWeld:斑马鱼短同源性诱导的高效靶向整合 作者:Jordan M. Welker, Wesley A. Wierson, Maira P. Almeida, Carla M. Mann, Melanie E. Torrie, Zhitao Ming, Stephen C. Ekker, Karl J. Clark, Drena L. Dobbs, Jeffrey J. Essner and Maura McGrail日期:07/20/2021,浏览量:110,Q&A: 0

Efficient precision genome engineering requires high frequency and specificity of integration at the genomic target site. Multiple design strategies for zebrafish gene targeting have previously been reported with widely varying frequencies for germline recovery of integration alleles. The GeneWeld protocol and pGTag (plasmids for Gene Tagging)

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免疫学

Isolation of Microglia and Analysis of Protein Expression by Flow Cytometry: Avoiding the Pitfall of Microglia Background Autofluorescence 小胶质细胞的分离及蛋白表达的流式细胞仪分析:避免小胶质细胞背景荧光的陷阱 作者:Jeremy C. Burns, Richard M. Ransohoff and Michaël Mingueneau日期:07/20/2021,浏览量:100,Q&A: 0

Microglia are a unique type of tissue-resident innate immune cell found within the brain, spinal cord, and retina. In the healthy nervous system, their main functions are to defend the tissue against infectious microbes, support neuronal networks through synapse remodeling, and clear extracellular debris and dying cells through phagocytosis. Many

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微生物学

In vitro Nitrate Reductase Activity Assay of Mycolicibacterium smegmatis Crude Extract 耻垢分枝杆菌粗提物硝酸还原酶活性的体外测定 作者:Wei Tan, Zhi-Hui Shao and Guo-Ping Zhao日期:07/20/2021,浏览量:78,Q&A: 0

Nitrate is one of the major inorganic nitrogen sources for microorganisms. Many bacterial and archaeal lineages can express cytoplasmic assimilatory nitrate reductase (NAS), which catalyzes the rate-limiting reduction of nitrate to nitrite in the nitrate assimilation pathway. Here, we present a detailed protocol for measuring in vitro nitrate

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分子生物学

Antisense Oligo Pulldown of Circular RNA for Downstream Analysis 用于下游分析的环状RNA的反义Oligo Pulldown 作者:Debojyoti Das, Aniruddha Das and Amaresh C. Panda日期:07/20/2021,浏览量:113,Q&A: 0

Circular RNAs (circRNAs) are a large family of noncoding RNA molecules that have emerged as novel regulators of gene expression by sequestering microRNAs (miRNAs) and RNA-binding proteins (RBPs). Several computational tools have been developed to predict circRNA interaction with target miRNAs and RBPs with a view to studying their potential effect

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A Simple Method to Generate Super-sensitive AID (ssAID)-based Conditional Knockouts using CRISPR-based Gene Knockout in Various Vertebrate Cell Lines 小胶质细胞的分离及蛋白表达的流式细胞仪分析:避免小胶质细胞背景荧光的陷阱 作者:Kohei Nishimura and Tatsuo Fukagawa日期:07/20/2021,浏览量:80,Q&A: 0

Inducing loss of function of a target protein using methods such as gene knockout is a powerful and useful strategy for analyzing protein function in cells. In recent years, the CRISPR/Cas-9-based gene knockout technology has been widely used across a variety of eukaryotes; however, this type of simple gene knockout strategy is not applicable to

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神经科学

Method for Rapid Enzymatic Cleaning for Reuse of Patch Clamp Pipettes: Increasing Throughput by Eliminating Manual Pipette Replacement between Patch Clamp Attempts 膜片钳移液管重复使用的快速酶清洗方法:通过消除膜片钳尝试之间的手动移液管更换来提高吞吐量 作者:Corey R. Landry, Mighten C. Yip, Ilya Kolb, William A. Stoy, Mercedes M. Gonzalez and Craig R. Forest日期:07/20/2021,浏览量:209,Q&A: 0

The whole-cell patch-clamp method is a gold standard for single-cell analysis of electrical activity, cellular morphology, and gene expression. Prior to our discovery that patch-clamp pipettes could be cleaned and reused, experimental throughput and automation were limited by the need to replace pipettes manually after each experiment. This

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植物科学

Spatiotemporal Quantification of Cytosolic pH in Arabidopsis Pollen Tubes 拟南芥花粉管胞质pH的时空定量研究 作者:Maria Teresa Portes, Daniel S.C. Damineli and José A. Feijó日期:07/20/2021,浏览量:149,Q&A: 0

Ion-specific probes and fluorescent indicators have been key in establishing the role of ion signaling, namely calcium, protons, and anions, in plant development, providing a robust approach for monitoring spatiotemporal changes in intracellular ion dynamics. The integration of protons/pH in signaling mechanisms is especially important as reports

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