生物化学

分类

    现刊
    Purification of Globular Actin from Rabbit Muscle and Pyrene Fluorescent Assays to Investigate Actin Dynamics in vitro
    从兔肌肉组织中纯化球状肌动蛋白并利用芘荧光分析实验体外检测肌动蛋白的动态变化
    作者:He Sun, Yuanyuan Luo and Yansong Miao日期:12/05/2018,浏览量:64,Q&A: 0
    Pyrene Fluorescent Assay is established to monitor the dynamic actin nucleation, elongation, capping and disassembly in vitro. This technique provides an easy handle procedure and straightforward visual data analysis. By coupling actin ...
    An in vitro Microscopy-based Assay for Microtubule-binding and Microtubule-crosslinking by Budding Yeast Microtubule-associated Protein
    利用芽殖酵母微管相关蛋白研究微管结合和微管交联的体外显微镜检测法
    作者:Yili Zhu, Weimin Tan and Wei-Lih Lee日期:12/05/2018,浏览量:169,Q&A: 0
    In this protocol, we describe a simple microscopy-based method to assess the interaction of a microtubule-associated protein (MAP) with microtubules. The interaction between MAP and microtubules is typically assessed by a co-sedimentation assay, ...
    Integrin Dependent RhoB Activation Assay Using Leukocytes
    白细胞内整合素依赖的RhoB激活实验
    作者:Janne Lehtonen, Katarzyna Potrzebowska, Malin Samuelsson and Lena Svensson日期:12/05/2018,浏览量:43,Q&A: 0
    To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the ...
    Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes
    流式细胞术检测T淋巴细胞中LFA-1的再循环
    作者:Katarzyna Potrzebowska, Janne Lehtonen, Malin Samuelsson and Lena Svensson日期:12/05/2018,浏览量:55,Q&A: 0
    To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type ...
    In vitro Membrane Interaction and Liposome Fusion Assays Using Recombinant Hepatitis C Virus Envelope Protein E2
    利用重组丙型肝炎病毒包膜蛋白E2体外检测膜互作和脂质体融合
    作者:Belén Yélamos, Mar Rodríguez-Rodríguez, Francisco Gavilanes and Julián Gómez-Gutiérrez日期:12/05/2018,浏览量:39,Q&A: 0
    In order to study the mechanism underlying the Hepatitis C Virus (HCV) fusion process we have performed assays using phospholipid liposomes and a truncated form of E2 protein, E2661 (amino acids 384-661 of the HCV polyprotein) lacking the ...
    Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis
    小麦种子凝集素(WGA)-SDS-PAGE:一种利用凝集素亲和凝胶电泳对O-乙酰氨基葡萄糖修饰蛋白进行检测的新方法
    作者:Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa日期:12/05/2018,浏览量:97,Q&A: 0
    Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, ...
    Purification of Globular Actin from Rabbit Muscle and Pyrene Fluorescent Assays to Investigate Actin Dynamics in vitro
    从兔肌肉组织中纯化球状肌动蛋白并利用芘荧光分析实验体外检测肌动蛋白的动态变化
    作者:He Sun, Yuanyuan Luo and Yansong Miao日期:12/05/2018,浏览量:64,Q&A: 0
    [Abstract] Pyrene Fluorescent Assay is established to monitor the dynamic actin nucleation, elongation, capping and disassembly in vitro. This technique provides an easy handle procedure and straightforward visual data analysis. By coupling actin purification and polymerization assays in this protocol, the readers could quickly get the affordable ...
    An in vitro Microscopy-based Assay for Microtubule-binding and Microtubule-crosslinking by Budding Yeast Microtubule-associated Protein
    利用芽殖酵母微管相关蛋白研究微管结合和微管交联的体外显微镜检测法
    作者:Yili Zhu, Weimin Tan and Wei-Lih Lee日期:12/05/2018,浏览量:169,Q&A: 0
    [Abstract] In this protocol, we describe a simple microscopy-based method to assess the interaction of a microtubule-associated protein (MAP) with microtubules. The interaction between MAP and microtubules is typically assessed by a co-sedimentation assay, which measures the amount of MAP that co-pellets with microtubules by centrifugation, followed by ...
    Integrin Dependent RhoB Activation Assay Using Leukocytes
    白细胞内整合素依赖的RhoB激活实验
    作者:Janne Lehtonen, Katarzyna Potrzebowska, Malin Samuelsson and Lena Svensson日期:12/05/2018,浏览量:43,Q&A: 0
    [Abstract] To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including ...
    Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes
    流式细胞术检测T淋巴细胞中LFA-1的再循环
    作者:Katarzyna Potrzebowska, Janne Lehtonen, Malin Samuelsson and Lena Svensson日期:12/05/2018,浏览量:55,Q&A: 0
    [Abstract] To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type in the human body, which express the leukocyte-specific integrin LFA-1. Here, we describe a flow ...
    In vitro Membrane Interaction and Liposome Fusion Assays Using Recombinant Hepatitis C Virus Envelope Protein E2
    利用重组丙型肝炎病毒包膜蛋白E2体外检测膜互作和脂质体融合
    作者:Belén Yélamos, Mar Rodríguez-Rodríguez, Francisco Gavilanes and Julián Gómez-Gutiérrez日期:12/05/2018,浏览量:39,Q&A: 0
    [Abstract] In order to study the mechanism underlying the Hepatitis C Virus (HCV) fusion process we have performed assays using phospholipid liposomes and a truncated form of E2 protein, E2661 (amino acids 384-661 of the HCV polyprotein) lacking the transmembrane region. E2661 has been previously generated by using the baculovirus ...
    Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis
    小麦种子凝集素(WGA)-SDS-PAGE:一种利用凝集素亲和凝胶电泳对O-乙酰氨基葡萄糖修饰蛋白进行检测的新方法
    作者:Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa日期:12/05/2018,浏览量:97,Q&A: 0
    [Abstract] Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, however, needs multiple and time-consuming steps if using conventional methods (e.g., ...
    Purification of Soluble Recombinant Human Tau Protein from Bacteria Using Double-tag Affinity Purification
    双标签亲和纯化法从细菌中纯化人可溶重组Tau蛋白
    作者:Joseph McInnes, Lujia Zhou and Patrik Verstreken日期:11/20/2018,浏览量:333,Q&A: 0
    [Abstract] Dysfunction of the microtubule-associated protein Tau (encoded by the MAPT gene) has been implicated in more than twenty neurodegenerative diseases, including Alzheimer’s. As such, the physiological and disease-relevant functions of Tau have garnered great interest in the research community. One barrier hampering investigations into the ...
    Relative Quantitation of Polymerized Actin in Suspension Cells by Flow Cytometry
    利用流式细胞术相对定量分析悬浮细胞中的聚合肌动蛋白
    作者:Mallory R. Kakley, Katrina B. Velle and Lillian K. Fritz-Laylin日期:11/20/2018,浏览量:350,Q&A: 0
    [Abstract] The amount of polymerized actin within a cell can vary widely due to natural processes and/or experimentally induced perturbations. We routinely use this protocol to measure relative polymerized actin content between cell populations by staining cells in suspension with fluorescent phalloidin, then measuring total cell fluorescence using flow ...
    Separation and Visualization of Low Abundant Ubiquitylated Forms
    低丰度泛素化形式蛋白的分离和可视化
    作者:Ramona Schuster, Tânia Simões, Fabian den Brave and Mafalda Escobar-Henriques日期:11/20/2018,浏览量:449,Q&A: 0
    [Abstract] In this protocol we describe the separation and visualization of ubiquitylated forms of the yeast mitofusin Fzo1 by Western blot. To this aim, we express HA-tagged Fzo1 in Saccharomyces cerevisiae, break the cells to extract a membrane-enriched fraction, solubilize the membranes using detergent and then specifically immunoprecipitate the ...
    Eicosanoid Isolation from Mouse Intestinal Tissue for ELISA
    从小鼠肠道组织中分离类二十烷酸用于ELISA
    作者:Isabella Rauch日期:11/05/2018,浏览量:283,Q&A: 0
    [Abstract] Activation of inflammasomes in peritoneal macrophages and intestinal epithelial cells (IEC) leads to the release of eicosanoids. To assess the amount of eicosanoids released by IEC, lipids need to be isolated from whole tissue previous to analysis by lipid mass spectrometry or ELISA. This protocol describes how to isolate lipids from intestinal ...