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生物化学

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    现刊
    In vitro Assays for Eukaryotic Leading/Lagging Strand DNA Replication
    真核先导链/后随链DNA复制的体外分析
    作者:Grant Schauer, Jeff Finkelstein and Mike O’Donnell日期:09/20/2017,浏览量:4,Q&A: 0
    The eukaryotic replisome is a multiprotein complex that duplicates DNA. The replisome is sculpted to couple continuous leading strand synthesis with discontinuous lagging strand synthesis, primarily carried out by DNA polymerases ε and δ, ...
    Isolation and Detection of the Chlorophyll Catabolite Hydroxylating Activity from Capsicum annuum Chromoplasts
    辣椒青霉色素中叶绿素的分离及含量羟化活性的测定
    作者:Mareike Hauenstein and Stefan Hörtensteiner日期:09/20/2017,浏览量:12,Q&A: 0
    Hydroxylation of chlorophyll catabolites at the so-called C32 position (Hauenstein et al., 2016) is commonly found in all plant species analyzed to date. Here we describe an in vitro hydroxylation assay using Capsicum ...
    Lipidomic Analysis of Caenorhabditis elegans Embryos
    秀丽隐杆线虫胚胎的脂质组学分析
    作者:Hung-Chi Yang, Cheng-Yu Hung, Yi-Yun Pan, Szecheng J Lo and Daniel Tsun-Yee Chiu日期:09/20/2017,浏览量:4,Q&A: 0
    Metabolomic is an emerging field of system biology. Lipidomic, a branch of metabolomic, aims to characterize lipophilic metabolites in biological systems. Caenorhabditis elegans (C. elegans) is a genetically tractable and versatile ...
    Protease Activity Assay in Fly Intestines
    苍蝇肠道蛋白酶活性测定
    作者:Marie-Paule Nawrot-Esposito, Rihab Loudhaief and Armel Gallet日期:09/20/2017,浏览量:12,Q&A: 0
    The intestine is a central organ required for the digestion of food, the absorption of nutrients and for fighting against aggressors ingested along with the food. Impairment of gut physiology following mucosal damages impacts its digestive ...
    Detection of Protein S-nitrosothiols (SNOs) in Plant Samples on Diaminofluorescein (DAF) Gels
    在二氨基荧光素(DAF)凝胶上检测植物样品中的蛋白质S-亚硝基硫醇(SNO)
    作者:Marta Rodríguez-Ruiz, Paulo T. Mioto, José M. Palma and Francisco J. Corpas 日期:09/20/2017,浏览量:10,Q&A: 0
    In plant cells, the analysis of protein S-nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for ...
    Arginine-rich Peptides Can Actively Mediate Liquid-liquid Phase Separation
    富含精氨酸的肽可以主动介导液-液相分离
    作者:Steven Boeynaems, Mathias De Decker, Peter Tompa and Ludo Van Den Bosch日期:09/05/2017,浏览量:130,Q&A: 0
    Studying liquid-liquid phase separation (LLPS) of proteins provides key insights into the biogenesis of membraneless organelles and pathological protein aggregation in disease. We have established a protocol for inducing the phase separation of ...
    Expression and Purification of a Mammalian P2X7 Receptor from Sf9 Insect Cells
    在Sf9昆虫细胞中表达纯化哺乳动物P2X7受体蛋白
    作者:Akira Karasawa and Toshimitsu Kawate日期:09/05/2017,浏览量:143,Q&A: 0
    The P2X7 receptor is an extracellular ATP-gated ion channel found only in eukaryotes (Bartlett et al., 2014). Due to its unique properties among P2X receptors, such as formation of a large conductance pore, the P2X7 receptor has been ...
    Labelling HaloTag Fusion Proteins with HaloTag Ligand in Living Cells
    在活细胞中使用HaloTag配体标记HaloTag融合蛋白
    作者:Huy Nguyen Duc and Xiaojun Ren 日期:09/05/2017,浏览量:70,Q&A: 0
    HaloTag has been widely used to label proteins in vitro and in vivo (Los et al., 2008). In this protocol, we describe labelling HaloTag-Cbx fusion proteins by HaloTag ligands for live-cell single-molecule imaging (Zhen et ...
    Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
    植物总提取物中泛素蛋白酶体活性测定
    作者:Suayib Üstün and Frederik Börnke日期:09/05/2017,浏览量:144,Q&A: 0
    The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through ...
    Extraction and Molybdenum Blue-based Quantification of Total Phosphate and Polyphosphate in Parachlorella
    拟小球藻中总磷酸盐和多磷酸盐的提取和钼蓝法定量
    作者:Shuhei Ota and Shigeyuki Kawano日期:09/05/2017,浏览量:73,Q&A: 0
    Inorganic phosphorus is a non-renewable resource and an essential element for life on Earth. Organisms such as algae, protists, and animals can store phosphate (Pi) through uptake of Pi as polyphosphate (poly-P), which is a linear polymer of ...
    Detection of Reactive Oxygen Species (ROS) in Cyanobacteria Using the Oxidant-sensing Probe 2’,7’-Dichlorodihydrofluorescein Diacetate (DCFH-DA)
    使用氧化物探针2',7'-二氯二氟荧光素二乙酸酯(DCFH-DA)检测蓝细菌中活性氧物质(ROS)
    作者:Rajneesh, Jainendra Pathak, Ananya Chatterjee, Shailendra P. Singh and Rajeshwar P. Sinha日期:09/05/2017,浏览量:111,Q&A: 0
    Reactive oxygen species (ROS) are cell signaling molecules synthesized inside the cells as a response to routine metabolic processes. In stress conditions such as ultraviolet radiation (UVR), ROS concentration increases several folds in the cells ...
    Sterol Analysis in Kluyveromyces lactis
    乳酸克鲁维酵母中的甾醇分析
    作者:Yvetta Gbelska, Nora Toth Hervay, Marcela Morvova and Alexandra Konecna日期:09/05/2017,浏览量:78,Q&A: 0
    Sterols are essential lipids of most eukaryotic cells with multiple functions (structural, regulatory and developmental). Sterol profile of yeast cells is often determined during the studies of ergosterol synthesis mutants used to uncover a number ...
    In vitro Assays for Eukaryotic Leading/Lagging Strand DNA Replication
    真核先导链/后随链DNA复制的体外分析
    作者:Grant Schauer, Jeff Finkelstein and Mike O’Donnell日期:09/20/2017,浏览量:4,Q&A: 0
    [Abstract] The eukaryotic replisome is a multiprotein complex that duplicates DNA. The replisome is sculpted to couple continuous leading strand synthesis with discontinuous lagging strand synthesis, primarily carried out by DNA polymerases ε and δ, respectively, along with helicases, polymerase α-primase, DNA sliding clamps, clamp loaders and many other ...
    Isolation and Detection of the Chlorophyll Catabolite Hydroxylating Activity from Capsicum annuum Chromoplasts
    辣椒青霉色素中叶绿素的分离及含量羟化活性的测定
    作者:Mareike Hauenstein and Stefan Hörtensteiner日期:09/20/2017,浏览量:12,Q&A: 0
    [Abstract] Hydroxylation of chlorophyll catabolites at the so-called C32 position (Hauenstein et al., 2016) is commonly found in all plant species analyzed to date. Here we describe an in vitro hydroxylation assay using Capsicum annuum chromoplast membranes as a source of the hydroxylating activity, which converts the ...
    Lipidomic Analysis of Caenorhabditis elegans Embryos
    秀丽隐杆线虫胚胎的脂质组学分析
    作者:Hung-Chi Yang, Cheng-Yu Hung, Yi-Yun Pan, Szecheng J Lo and Daniel Tsun-Yee Chiu日期:09/20/2017,浏览量:4,Q&A: 0
    [Abstract] Metabolomic is an emerging field of system biology. Lipidomic, a branch of metabolomic, aims to characterize lipophilic metabolites in biological systems. Caenorhabditis elegans (C. elegans) is a genetically tractable and versatile animal model for novel discovery of lipid metabolism. In addition, C. elegans embryo is ...
    Protease Activity Assay in Fly Intestines
    苍蝇肠道蛋白酶活性测定
    作者:Marie-Paule Nawrot-Esposito, Rihab Loudhaief and Armel Gallet日期:09/20/2017,浏览量:12,Q&A: 0
    [Abstract] The intestine is a central organ required for the digestion of food, the absorption of nutrients and for fighting against aggressors ingested along with the food. Impairment of gut physiology following mucosal damages impacts its digestive capacities that consequently will affect growth, wellbeing or even survival of the individual. Hence, the ...
    Detection of Protein S-nitrosothiols (SNOs) in Plant Samples on Diaminofluorescein (DAF) Gels
    在二氨基荧光素(DAF)凝胶上检测植物样品中的蛋白质S-亚硝基硫醇(SNO)
    作者:Marta Rodríguez-Ruiz, Paulo T. Mioto, José M. Palma and Francisco J. Corpas 日期:09/20/2017,浏览量:10,Q&A: 0
    [Abstract] In plant cells, the analysis of protein S-nitrosothiols (SNOs) under physiological and adverse stress conditions is essential to understand the mechanisms of Nitric oxide (NO)-based signaling. We adapted a previously reported protocol for detecting protein SNOs in animal systems (King et al., 2005) for plant samples. Briefly, ...
    Arginine-rich Peptides Can Actively Mediate Liquid-liquid Phase Separation
    富含精氨酸的肽可以主动介导液-液相分离
    作者:Steven Boeynaems, Mathias De Decker, Peter Tompa and Ludo Van Den Bosch日期:09/05/2017,浏览量:130,Q&A: 0
    [Abstract] Studying liquid-liquid phase separation (LLPS) of proteins provides key insights into the biogenesis of membraneless organelles and pathological protein aggregation in disease. We have established a protocol for inducing the phase separation of arginine-rich peptides, which allows for studying their molecular determinants and dynamics (Boeynaems ...
    Expression and Purification of a Mammalian P2X7 Receptor from Sf9 Insect Cells
    在Sf9昆虫细胞中表达纯化哺乳动物P2X7受体蛋白
    作者:Akira Karasawa and Toshimitsu Kawate日期:09/05/2017,浏览量:143,Q&A: 0
    [Abstract] The P2X7 receptor is an extracellular ATP-gated ion channel found only in eukaryotes (Bartlett et al., 2014). Due to its unique properties among P2X receptors, such as formation of a large conductance pore, the P2X7 receptor has been implicated in devastating diseases like chronic pain (North and Jarvis, 2013). However, mechanisms ...
    Labelling HaloTag Fusion Proteins with HaloTag Ligand in Living Cells
    在活细胞中使用HaloTag配体标记HaloTag融合蛋白
    作者:Huy Nguyen Duc and Xiaojun Ren 日期:09/05/2017,浏览量:70,Q&A: 0
    [Abstract] HaloTag has been widely used to label proteins in vitro and in vivo (Los et al., 2008). In this protocol, we describe labelling HaloTag-Cbx fusion proteins by HaloTag ligands for live-cell single-molecule imaging (Zhen et al., 2016).
    Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
    植物总提取物中泛素蛋白酶体活性测定
    作者:Suayib Üstün and Frederik Börnke日期:09/05/2017,浏览量:144,Q&A: 0
    [Abstract] The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to ...
    Extraction and Molybdenum Blue-based Quantification of Total Phosphate and Polyphosphate in Parachlorella
    拟小球藻中总磷酸盐和多磷酸盐的提取和钼蓝法定量
    作者:Shuhei Ota and Shigeyuki Kawano日期:09/05/2017,浏览量:73,Q&A: 0
    [Abstract] Inorganic phosphorus is a non-renewable resource and an essential element for life on Earth. Organisms such as algae, protists, and animals can store phosphate (Pi) through uptake of Pi as polyphosphate (poly-P), which is a linear polymer of orthophosphate residues linked by high-energy phosphoanhydride bonds. Here, we describe procedures for ...