• 卷册 11, 2021
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  • 卷册 9, 2019
  • 卷册 8, 2018
  • 卷册 7, 2017
  • 卷册 6, 2016
  • 卷册 5, 2015
  • 卷册 4, 2014
  • 卷册 3, 2013
  • 卷册 2, 2012
  • 卷册 1, 2011
Image Credit: BioProtoc.4127

往期刊物(2021)

卷册: 11 期号: 16

生物化学

Monitoring Protein Splicing Using In-gel Fluorescence Immediately Following SDS-PAGE SDS-PAGE后立即使用凝胶内荧光监测蛋白质剪接 作者:Joel Weinberger II and Christopher W. Lennon日期:08/20/2021,浏览量:1168,Q&A: 0

Inteins garner significant interest from both basic and applied researchers due to their unique catalytic abilities. Herein, we describe a protocol for accurately monitoring protein splicing without purification using in-gel fluorescence immediately following Tris-Glycine SDS-PAGE. Following expression in Escherichia coli, cells are lysed by

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Oil Red O Staining for Lipid Content in Caenorhabditis elegans 油红O染色测定秀丽隐杆线虫脂质含量   作者:Feng-Yung Wang and Tsui-Ting Ching日期:08/20/2021,浏览量:885,Q&A: 0

The nematode Caenorhabditis elegans has emerged as a popular model system for studying the regulation of lipid metabolism. Therefore, it is critical to develop a method for determining fat storage in individual worms. Oil Red O (ORO) staining has been validated as an accurate assessment for major fat storage in C. elegans. Here, we describe an

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Implementing Novel Designs in pET Expression Plasmids that Increase Protein Production 在pET表达质粒中实现增加蛋白质产量的新设计 作者:Patrick J. Shilling and Daniel O. Daley日期:08/20/2021,浏览量:1344,Q&A: 0

pET expression plasmids are widely used in the biotechnology, biopharmaceutical, and basic research sectors for the production of recombinant proteins. Typically, they are used off-the-shelf because they support high production titers; however, we have identified two design flaws in many pET plasmids that limit their production capacity. We used

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生物工程

3D-printed Recoverable Microdrive and Base Plate System for Rodent Electrophysiology 用于啮齿动物电生理学的3D打印可恢复微驱动器和基板系统 作者:Mihály Vöröslakos, Hiroyuki Miyawaki, Sebastien Royer, Kamran Diba, Euisik Yoon, Peter C. Petersen and György Buzsáki日期:08/20/2021,浏览量:863,Q&A: 0

Extracellular recordings in freely moving animals allow the monitoring of brain activity from populations of neurons at single-spike temporal resolution. While state-of-the-art electrophysiological recording devices have been developed in recent years (e.g., µLED and Neuropixels silicon probes), implantation methods for silicon probes in

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生物物理学

Unraveling the Physicochemical Determinants of Protein Liquid-liquid Phase Separation by Nanoscale Infrared Vibrational Spectroscopy 利用纳米尺度红外振动光谱揭示蛋白质液-液相分离的理化决定因素 作者:Francesco S. Ruggeri, Alyssa M. Miller, Michele Vendruscolo and Tuomas P. J. Knowles日期:08/20/2021,浏览量:906,Q&A: 0

The phenomenon of reversible liquid-liquid phase separation of proteins underlies the formation of membraneless organelles, which are crucial for cellular processes such as signalling and transport. In addition, it is also of great interest to uncover the mechanisms of further irreversible maturation of the functional dense liquid phase into

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Synchronized Real-time Measurement of Sec-mediated Protein Translocation Sec介导的蛋白质易位的同步实时测量 作者:Riti Gupta, Dmitri Toptygin and Christian M. Kaiser日期:08/20/2021,浏览量:679,Q&A: 0

The Sec translocon, consisting of a heterotrimeric transmembrane channel (SecYEG) and an associated ATPase (SecA), catalyzes the export of unfolded proteins from the cytosol in bacteria. Kinetically resolving protein translocation at high resolution yields mechanistic insight into the process. Translocation is typically followed by measuring the

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癌症生物学

Measuring DNA Damage Using the Alkaline Comet Assay in Cultured Cells 培养细胞DNA损伤的碱性彗星试验 作者:Elena Clementi, Zuzana Garajova and Enni Markkanen日期:08/20/2021,浏览量:958,Q&A: 0

Maintenance of DNA integrity is of pivotal importance for cells to circumvent detrimental processes that can ultimately lead to the development of various diseases. In the face of a plethora of endogenous and exogenous DNA-damaging agents, cells have evolved a variety of DNA repair mechanisms that are responsible for safeguarding genetic

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细胞生物学

A Novel Method to Make Polyacrylamide Gels with Mechanical Properties Resembling those of Biological Tissues 一种制备具有类似生物组织力学性能的聚丙烯酰胺凝胶的新方法   作者:Katarzyna Pogoda, Elisabeth E. Charrier and Paul A. Janmey日期:08/20/2021,浏览量:946,Q&A: 0

Studies characterizing how cells respond to the mechanical properties of their environment have been enabled by the use of soft elastomers and hydrogels as substrates for cell culture. A limitation of most such substrates is that, although their elastic properties can be accurately controlled, their viscous properties cannot, and cells

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发育生物学

Preparation of Drosophila Larval Blood Cells for Single-cell RNA Sequencing 果蝇幼虫血细胞单细胞RNA测序的制备 作者:Sudhir Gopal Tattikota and Norbert Perrimon日期:08/20/2021,浏览量:530,Q&A: 0

Recent advances in single-cell RNA-sequencing (scRNA-seq) technologies provide unprecedented opportunities to identify new cell types and characterize cell states. One of the most important requirements for performing scRNA-seq is to obtain high-quality single cells in suspension. Recently, we used this approach to characterize Drosophila blood

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免疫学

Construction of a Highly Diverse mRNA Library for in vitro Selection of Monobodies 体外选择单体的高度多样性mRNA文库的构建 作者:Taishi Kondo, Minori Eguchi, Nariaki Tsuzuki, Naoya Murata, Tomoshige Fujino, Gosuke Hayashi and Hiroshi Murakami日期:08/20/2021,浏览量:530,Q&A: 0

Recently, we developed transcription/translation coupled with the association of puromycin linker (TRAP) display as a quick in vitro selection method to obtain antibody-like proteins. For the in vitro selection, it is important to prepare mRNA libraries among which the diversity is high. Here, we describe a method for the preparation of monobody

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One-step White Blood Cell Extracellular Staining Method for Flow Cytometry 流式细胞术白细胞细胞外一步染色法 作者:Ines Ait Belkacem, Pénélope Bourgoin, Jean Marc Busnel, Franck Galland and Fabrice Malergue日期:08/20/2021,浏览量:682,Q&A: 0

Flow cytometry is a powerful analytical technique that is increasingly used in scientific investigations and healthcare; however, it requires time-consuming, multi-step sample procedures, which limits its use to specialized laboratories. In this study, we propose a new universal one-step method in which white blood cell staining and red blood cell

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微生物学

Acid Hydrolysis for the Extraction of Archaeal Core Lipids and HPLC-MS Analysis 酸水解法提取古生菌核心脂质及HPLC-MS分析 作者:Maxime Tourte, Philippe Schaeffer, Vincent Grossi and Philippe M. Oger日期:08/20/2021,浏览量:387,Q&A: 0

Lipid membranes are essential cellular elements as they provide cellular integrity and selective permeability under a broad range of environmental settings upon cell growth. In particular, Archaea are commonly recognized for their tolerance to extreme conditions, which is now widely accepted to stem from the unique structure of their lipids. While

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Development and Quantitation of Pseudomonas aeruginosa Biofilms after in vitro Cultivation in Flow-reactors 流动反应器体外培养铜绿假单胞菌生物膜的研制与定量 作者:Yingdan Zhang, Jingru Zhao, Hang Cheng, Jing Wang, Liang Yang and Haihua Liang日期:08/20/2021,浏览量:379,Q&A: 0

Characterization of biofilm formation and metabolic activities is critical to investigating biofilm interactions with environmental factors and illustrating biofilm regulatory mechanisms. An appropriate in vitro model that mimics biofilm in vivo habitats therefore demands accurate quantitation and investigation of biofilm-associated activities.

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分子生物学

Optimised Method for the Production and Titration of Lentiviral Vectors Pseudotyped with the SARS-CoV-2 Spike SARS-CoV-2刺突慢病毒伪型载体制备和滴定的优化方法 作者:Leila Mekkaoui, Emma M. Bentley, Mathieu Ferrari, Katarina Lamb, Katarzyna Ward, Rajeev Karattil, Zulaikha Akbar, Reyisa Bughda, James Sillibourne, Shimobi Onuoha, Giada Mattiuzzo, Yasu Takeuchi and Martin Pule日期:08/20/2021,浏览量:400,Q&A: 0

The use of recombinant lentivirus pseudotyped with the coronavirus Spike protein of SARS-CoV-2 would circumvent the requirement of biosafety-level 3 (BSL-3) containment facilities for the handling of SARS-CoV-2 viruses. Herein, we describe a fast and reliable protocol for the transient production of lentiviruses pseudotyped with SARS-CoV-2 Spike

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植物科学

A Novel Method to Map Small RNAs with High Resolution 一种新的高分辨率小RNA图谱绘制方法 作者:Kun Huang, Feray Demirci, Blake C. Meyers and Jeffrey L. Caplan日期:08/20/2021,浏览量:614,Q&A: 0

Analyzing cellular structures and the relative location of molecules is essential for addressing biological questions. Super-resolution microscopy techniques that bypass the light diffraction limit have become increasingly popular to study cellular molecule dynamics in situ. However, the application of super-resolution imaging techniques to detect

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Tomato Stem Injection for the Precise Assessment of Ralstonia solanacearum Fitness in Planta 番茄茎杆注射法精确评价青枯菌在植物中的适应性 作者:Yaru Wang, Liu Xian, Gang Yu and Alberto P. Macho日期:08/20/2021,浏览量:438,Q&A: 0

Ralstonia solanacearum is a soil-borne pathogen with worldwide distribution that causes bacterial wilt disease in more than 250 plant species. R. solanacearum invades plants through the roots, reaches the vascular system, and colonizes the whole plant by moving through the xylem, where it eventually replicates rapidly, causing plant death. Usual

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干细胞

Isolation of Single Cells from Mouse Periodontal Ligament 小鼠牙周膜单细胞的分离 作者:Jing Zhao and Paul Sharpe日期:08/20/2021,浏览量:892,Q&A: 0

The periodontal ligament (PDL) is an essential tissue connecting teeth and bone. It is a complex tissue specifically designed to absorb the forces of mastication; analysis of its multiple cell populations is important to understand its function and the cell changes associated with periodontal disease. Cells in the periodontal ligament are not

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Neutral Comet Assay to Detect and Quantitate DNA Double-Strand Breaks in Hematopoietic Stem Cells 检测和定量造血干细胞DNA双链断裂的中性彗星试验 作者:Irene Mariam Roy, Pon Sowbhagya Nadar and Satish Khurana日期:08/20/2021,浏览量:524,Q&A: 0

In vertebrates, hematopoietic stem cells (HSCs) regulate the supply of blood cells throughout the lifetime and help to maintain homeostasis. Due to their long lifespan, genetic integrity is paramount for these cells, and accordingly, a number of stem cell-specific mechanisms are employed. However, HSCs tend to show more DNA damage with increasing

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