生物化学

分类

    现刊
    FRET-based Microscopy Assay to Measure Activity of Membrane Amino Acid Transporters with Single-transporter Resolution
    用荧光共振能量转移的显微技术测定单转运体膜氨基酸转运体的活性
    [Abstract]

    Secondary active transporters reside in cell membranes transporting polar solutes like amino acids against steep concentration gradients, using electrochemical gradients of ions as energy sources. Commonly, ensemble-based measurements of radiolabeled substrate uptakes or transport currents inform on kinetic parameters of transporters. Here we

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    Conjugation of Fab’ Fragments with Fluorescent Dyes for Single-molecule Tracking on Live Cells
    Fab片段结合荧光染料用于活细胞单分子示踪
    作者:I-Ting Teng, Xiangning Bu and Inhee Chung日期:09/20/2019,浏览量:3369,Q&A: 0
    [Abstract] Our understanding of the regulation and functions of cell-surface proteins has progressed rapidly with the advent of advanced optical imaging techniques. In particular, single-molecule tracking (SMT) using bright fluorophores conjugated to antibodies and wide-field microscopy methods such as total internal reflection fluorescence microscopy have ...
    Preparation of Cell-free Synthesized Proteins Selectively Double Labeled for Single-molecule FRET Studies
    用于单分子FRET研究的选择性双标记无细胞合成蛋白质的制备
    作者:Mayuri Sadoine, Michele Cerminara, Jörg Fitter and Alexandros Katranidis日期:06/20/2018,浏览量:4072,Q&A: 0
    [Abstract] Single-molecule FRET (smFRET) is a powerful tool to investigate molecular structures and conformational changes of biological molecules. The technique requires protein samples that are site-specifically equipped with a pair of donor and acceptor fluorophores. Here, we present a detailed protocol for preparing double-labeled proteins for smFRET ...
    Fluorophore Labeling, Nanodisc Reconstitution and Single-molecule Observation of a G Protein-coupled Receptor
    G蛋白偶联受体的荧光标记、磷脂纳米盘重建和单分子观察
    作者:Rajan Lamichhane, Jeffrey J. Liu, Raymond F. Pauszek III and David P. Millar日期:06/20/2017,浏览量:6851,Q&A: 0
    [Abstract] Activation of G protein-coupled receptors (GPCRs) by agonist ligands is mediated by a transition from an inactive to active receptor conformation. We describe a novel single-molecule assay that monitors activation-linked conformational transitions in individual GPCR molecules in real-time. The receptor is site-specifically labeled with a Cy3 ...
    Cloud-point PEG Glass Surfaces for Imaging of Immobilized Single Molecules by Total-internal-reflection Microscopy
    采用浊点PEG玻璃表面法和全内反射显微镜进行固定化单分子成像
    作者:Zhengjian Zhang, Sang Ryul Park, Alexandros Pertsinidis and Andrey Revyakin日期:04/05/2016,浏览量:8948,Q&A: 0
    [Abstract] This effective, robust protocol generates glass coverslips coated with biotin-functionalized polyethylene glycol (PEG), making the glass surface resistant to non-specific absorption of biomolecules, and permitting immobilization of biomolecules for subsequent single-molecule tracking of biochemical reactions. The protocol can be completed in one ...