生物化学

分类

    现刊
    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:1149,Q&A: 0
    [Abstract]

    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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    A New Method for Studying RNA-binding Proteins on Specific RNAs
    一种研究特异性RNA结合蛋白的新方法
    作者:Weiping Sun, Ziheng Zhang, Ji-Long Liu and Min Zhuang日期:05/20/2021,浏览量:6360,Q&A: 0
    [Abstract]

    Proximity-based protein labeling has been developed to identify protein-nucleic acid interactions. We have reported a novel method termed CRUIS (CRISPR-based RNA-United Interacting System), which captures RNA-protein interactions in living cells by combining the RNA-binding capacity of CRISPR/Cas13 and the proximity-tagging activity of PUP-IT.

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    In vitro Induction and Detection of Acrosomal Exocytosis in Human Spermatozoa
    人精子中顶体胞吐作用的体外诱导和检测
    作者:Shenae L. Cafe, Amanda L. Anderson and Brett Nixon日期:07/20/2020,浏览量:1776,Q&A: 0
    [Abstract] The acrosome reaction is a highly regulated exocytotic event that primes spermatozoa for successful fertilization. Upon induction, acrosomal exocytosis proceeds via a wave of vesiculation that radiates across the sperm head, destabilizing the acrosomal vesicle and resulting in the release of the acrosomal contents. Having shed their acrosome, ...
    Imaging VIPER-labeled Cellular Proteins by Correlative Light and Electron Microscopy
    利用关联光学和电子显微镜技术进行VIPER标记细胞蛋白成像
    作者:Julia K. Doh, Young Hwan Chang, Caroline A. Enns, Claudia S. Lόpez and Kimberly E. Beatty日期:11/05/2019,浏览量:3235,Q&A: 0
    [Abstract] Advances in fluorescence microscopy (FM), electron microscopy (EM), and correlative light and EM (CLEM) offer unprecedented opportunities for studying diverse proteins and nanostructures involved in fundamental cell biology. It is now possible to visualize and quantify the spatial organization of cellular proteins and other macromolecules by FM, ...
    Implementing VIPER for Imaging Cellular Proteins by Fluorescence Microscopy
    VIPER用于荧光显微镜中细胞蛋白成像
    作者:Julia K. Doh, Caroline A. Enns and Kimberly E. Beatty日期:11/05/2019,浏览量:2891,Q&A: 0
    [Abstract] Genetically-encoded tags are useful tools for multicolor and multi-scale cellular imaging. Versatile Interacting Peptide (VIP) tags, such as VIPER, are new genetically-encoded tags that can be used in various imaging applications. VIP tags consist of a coiled-coil heterodimer, with one peptide serving as the genetic tag and the other (“probe ...
    Generation of CoilR Probe Peptides for VIPER-labeling of Cellular Proteins
    用于细胞蛋白VIPER标记的CoilR肽探针的制备
    作者:Julia K. Doh, Savannah J. Tobin and Kimberly E. Beatty日期:11/05/2019,浏览量:2785,Q&A: 0
    [Abstract] Versatile Interacting Peptide (VIP) tags are a new class of genetically-encoded tag designed for imaging cellular proteins by fluorescence and electron microscopy. In 2018, we reported the VIPER tag (Doh et al., 2018), which contains two elements: a genetically-encoded peptide tag (i.e., CoilE) and a probe peptide (i.e., ...
    Photoaffinity Labeling of Respiratory Complex I in Bovine Heart Submitochondrial Particles by Photoreactive [125I] amilorides
    利用光敏阿米洛利光亲和标记牛心脏亚线粒体颗粒中呼吸链复合物I
    作者:Masatoshi Murai and Hideto Miyoshi日期:09/05/2019,浏览量:2605,Q&A: 0
    [Abstract] The architecture of quinone/inhibitor-access channel in proton-translocating NADH-quinone oxidoreductase (respiratory complex I) was modeled by X-ray crystallography and cryo-EM, however, it remains debatable whether the channel model reflects the physiologically relevant state present throughout the catalytic cycle. Using photoreactive [125 ...
    Click Chemistry (CuAAC) and Detection of Tagged de novo Synthesized Proteins in Drosophila
    果蝇中通过“链接”化学检测标记的从头合成蛋白
    [Abstract] Copper-catalyzed azide-alkyne-cycloaddition (CuAAC), also known as ‘click chemistry’ serves as a technique for bio-orthogonal, that is, bio-compatible labeling of macromolecules including proteins or lipids. Click chemistry has been widely used to covalently, selectively, and efficiently attach probes such as fluorophores or biotin to small ...
    Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging
    用于3D荧光成像的鼠尾胶的荧光标记
    作者:Andrew D. Doyle日期:07/05/2018,浏览量:6229,Q&A: 0
    [Abstract] Rat tail collagen solutions have been used as polymerizable in vitro three-dimensional (3D) extracellular matrix (ECM) gels for single and collective cell migration assays as well as spheroid formation. These 3D hydrogels are a relatively inexpensive, simple to use model system that can mimic the in vivo physical characteristics ...
    A General Method for Intracellular Protein Delivery through ‘E-tag’ Protein Engineering and Arginine Functionalized Gold Nanoparticles
    通过'E标签'蛋白质工程和精氨酸功能化金纳米粒子进行细胞内蛋白质递送的通用方法
    作者:Rubul Mout and Vincent M. Rotello日期:12/20/2017,浏览量:5899,Q&A: 0
    [Abstract] In this protocol, we describe a method for direct cytosolic protein delivery that avoids endosomal entrapment of the delivered proteins. We achieved this by tagging the desired protein with an oligo glutamic acid tag (E-tag), and subsequently using carrier gold nanoparticles to deliver these E-tagged proteins. When E-tagged proteins and ...