哺乳纲

分类

    现刊
    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:19,Q&A: 0
    [Abstract]

    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:17,Q&A: 0
    [Abstract]

    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 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:16,Q&A: 0
    [Abstract]

    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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    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:20,Q&A: 0
    [Abstract]

    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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    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:13,Q&A: 0
    [Abstract]

    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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    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:556,Q&A: 0
    [Abstract]

    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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    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:808,Q&A: 0
    [Abstract]

    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:818,Q&A: 0
    [Abstract]

    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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    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:432,Q&A: 0
    [Abstract]

    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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    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:856,Q&A: 0
    [Abstract]

    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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