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生物化学

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    现刊
    Detection of Intracellular Reduced (Catalytically Active) SHP-1 and Analyses of Catalytically Inactive SHP-1 after Oxidation by Pervanadate or H2O2
    细胞内还原的(具有催化活性的)SHP-1检测以及在经钒酸盐或H2O2氧化后无催化活性SHP-1的分析
    作者:Seeyoung Choi and Paul E. Love日期:01/05/2018,浏览量:174,Q&A: 0
    Oxidative inactivation of cysteine-dependent Protein Tyrosine Phosphatases (PTPs) by cellular reactive oxygen species (ROS) plays a critical role in regulating signal transduction in multiple cell types. The phosphatase activity of most PTPs depends ...
    In vitro RNA-dependent RNA Polymerase Assay Using Arabidopsis RDR6
    拟南芥RDR6的体外RNA依赖性RNA聚合酶活性测定
    作者:Kyungmin Baeg, Yukihide Tomari and Hiro-oki Iwakawa日期:01/05/2018,浏览量:117,Q&A: 0
    RNA-dependent RNA polymerases (RdRPs) in eukaryotes convert single-stranded RNAs into double-stranded RNAs, thereby amplifying small interfering RNAs that play crucial roles in the regulation of development, maintenance of genome integrity and ...
    High Throughput NPY-Venus and Serotonin Secretion Assays for Regulated Exocytosis in Neuroendocrine Cells
    高通量NPY-Venus和5-羟色胺分泌实验分析神经内分泌细胞中被调控的胞吐作用
    作者:Xingmin Aaron Zhang and Thomas F.J. Martin日期:01/05/2018,浏览量:153,Q&A: 0
    Here we describe two assays to measure dense core vesicle (DCV) exocytosis-mediated cargo secretion in neuroendocrine cells. To conduct siRNA screens for novel genes in regulated DCV exocytosis, we developed a plate reader-based secretion assay ...
    Determining Ribosome Translational Status by Ribo-ELISA
    采用Ribo-ELISA检测核糖体的翻译状态
    作者:Amandine Bastide, Jonathan W. Yewdell and Alexandre David日期:01/05/2018,浏览量:258,Q&A: 0
    The Ribo-ELISA was originally developed to elucidate the basis for the ribopuromycylation method (RPM)-based detection of ribosome bound nascent chains. The Ribo-ELISA enables characterization of the translational status of ribosomes, and can be ...
    Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons
    Neuro2A细胞和大鼠原代皮层神经元中突变亨廷顿蛋白的分泌
    作者:Katarina Trajkovic, Hyunkyung Jeong and Dimitri Krainc日期:01/05/2018,浏览量:111,Q&A: 0
    Quantitative analysis of proteins secreted from the cells poses a challenge due to their low abundance and the interfering presence of a large amount of bovine serum albumin (BSA) in the cell culture media. We established assays for detection of ...
    Analysis of Direct Interaction between Viral DNA-binding Proteins by Protein Pull-down Co-immunoprecipitation Assay
    蛋白质pull-down免疫共沉淀实验分析病毒DNA结合蛋白质之间的直接相互作用
    作者:Ana Lechuga, Mónica Berjón-Otero, Margarita Salas and Modesto Redrejo-Rodríguez日期:01/05/2018,浏览量:221,Q&A: 0
    This protocol analyzes the direct interaction between two DNA-binding proteins by pull-down co-immunoprecipitation. One of the proteins is overexpressed in E. coli as HA-tagged recombinant protein and cell-free extracts are ...
    Visible Immunoprecipitation (VIP) Assay: a Simple and Versatile Method for Visual Detection of Protein-protein Interactions
    可视免疫沉淀(VIP)实验:一种简单通用的蛋白质 - 蛋白质相互作用可视化检测方法
    作者:Yohei Katoh, Kentaro Nakamura and Kazuhisa Nakayama日期:01/05/2018,浏览量:232,Q&A: 0
    The visible immunoprecipitation (VIP) assay is a convenient alternative to conventional co-immunoprecipitation (Katoh et al., 2015). By processing lysates from cells co-expressing GFP-fusion and RFP-fusion proteins for immunoprecipitation ...
    Fatty Acid Content and Composition of Triacylglycerols of Chlorella kessleri
    凯氏小球藻中脂肪酸含量及三酰甘油组成
    作者:Motohide Aoki and Norihiro Sato日期:01/05/2018,浏览量:110,Q&A: 0
    Triacylglycerols (TAGs) are esters formed from one glycerol and three fatty acids. TAGs are induced to accumulate in algal cells under environmental stress conditions including nutrient-limitation, hyperosmosis, and low temperature, for the storage ...
    Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER
    使用B-PER简单有效地透化蓝藻以进行体内酶分析
    作者:Simon Matthé Erstad and Yumiko Sakuragi日期:01/05/2018,浏览量:172,Q&A: 0
    Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels ...
    Detection of Intracellular Reduced (Catalytically Active) SHP-1 and Analyses of Catalytically Inactive SHP-1 after Oxidation by Pervanadate or H2O2
    细胞内还原的(具有催化活性的)SHP-1检测以及在经钒酸盐或H2O2氧化后无催化活性SHP-1的分析
    作者:Seeyoung Choi and Paul E. Love日期:01/05/2018,浏览量:174,Q&A: 0
    [Abstract] Oxidative inactivation of cysteine-dependent Protein Tyrosine Phosphatases (PTPs) by cellular reactive oxygen species (ROS) plays a critical role in regulating signal transduction in multiple cell types. The phosphatase activity of most PTPs depends upon a ‘signature’ cysteine residue within the catalytic domain that is maintained in the ...
    In vitro RNA-dependent RNA Polymerase Assay Using Arabidopsis RDR6
    拟南芥RDR6的体外RNA依赖性RNA聚合酶活性测定
    作者:Kyungmin Baeg, Yukihide Tomari and Hiro-oki Iwakawa日期:01/05/2018,浏览量:117,Q&A: 0
    [Abstract] RNA-dependent RNA polymerases (RdRPs) in eukaryotes convert single-stranded RNAs into double-stranded RNAs, thereby amplifying small interfering RNAs that play crucial roles in the regulation of development, maintenance of genome integrity and antiviral immunity. Here, we describe a method of in vitro RdRP assay using recombinant ...
    High Throughput NPY-Venus and Serotonin Secretion Assays for Regulated Exocytosis in Neuroendocrine Cells
    高通量NPY-Venus和5-羟色胺分泌实验分析神经内分泌细胞中被调控的胞吐作用
    作者:Xingmin Aaron Zhang and Thomas F.J. Martin日期:01/05/2018,浏览量:153,Q&A: 0
    [Abstract] Here we describe two assays to measure dense core vesicle (DCV) exocytosis-mediated cargo secretion in neuroendocrine cells. To conduct siRNA screens for novel genes in regulated DCV exocytosis, we developed a plate reader-based secretion assay using DCV cargo, NPY-Venus, and an orthogonal 3H-serotonin secretion assay. The NPY-Venus ...
    Determining Ribosome Translational Status by Ribo-ELISA
    采用Ribo-ELISA检测核糖体的翻译状态
    作者:Amandine Bastide, Jonathan W. Yewdell and Alexandre David日期:01/05/2018,浏览量:258,Q&A: 0
    [Abstract] The Ribo-ELISA was originally developed to elucidate the basis for the ribopuromycylation method (RPM)-based detection of ribosome bound nascent chains. The Ribo-ELISA enables characterization of the translational status of ribosomes, and can be applied to the discovery of super-ribosomal complexes with novel ribosome associated macromolecules ...
    Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons
    Neuro2A细胞和大鼠原代皮层神经元中突变亨廷顿蛋白的分泌
    作者:Katarina Trajkovic, Hyunkyung Jeong and Dimitri Krainc日期:01/05/2018,浏览量:111,Q&A: 0
    [Abstract] Quantitative analysis of proteins secreted from the cells poses a challenge due to their low abundance and the interfering presence of a large amount of bovine serum albumin (BSA) in the cell culture media. We established assays for detection of mutant huntingtin (mHtt) secreted from Neuro2A cell line stably expressing mHtt and rat primary ...
    Analysis of Direct Interaction between Viral DNA-binding Proteins by Protein Pull-down Co-immunoprecipitation Assay
    蛋白质pull-down免疫共沉淀实验分析病毒DNA结合蛋白质之间的直接相互作用
    作者:Ana Lechuga, Mónica Berjón-Otero, Margarita Salas and Modesto Redrejo-Rodríguez日期:01/05/2018,浏览量:221,Q&A: 0
    [Abstract] This protocol analyzes the direct interaction between two DNA-binding proteins by pull-down co-immunoprecipitation. One of the proteins is overexpressed in E. coli as HA-tagged recombinant protein and cell-free extracts are immunoprecipitated in HA-affinity resin. Cell extracts are treated with nuclease to degrade DNA and RNA, which rules ...
    Visible Immunoprecipitation (VIP) Assay: a Simple and Versatile Method for Visual Detection of Protein-protein Interactions
    可视免疫沉淀(VIP)实验:一种简单通用的蛋白质 - 蛋白质相互作用可视化检测方法
    作者:Yohei Katoh, Kentaro Nakamura and Kazuhisa Nakayama日期:01/05/2018,浏览量:232,Q&A: 0
    [Abstract] The visible immunoprecipitation (VIP) assay is a convenient alternative to conventional co-immunoprecipitation (Katoh et al., 2015). By processing lysates from cells co-expressing GFP-fusion and RFP-fusion proteins for immunoprecipitation with GST-tagged anti-GFP Nanobody and glutathione-Sepharose beads, protein-protein interactions can ...
    Fatty Acid Content and Composition of Triacylglycerols of Chlorella kessleri
    凯氏小球藻中脂肪酸含量及三酰甘油组成
    作者:Motohide Aoki and Norihiro Sato日期:01/05/2018,浏览量:110,Q&A: 0
    [Abstract] Triacylglycerols (TAGs) are esters formed from one glycerol and three fatty acids. TAGs are induced to accumulate in algal cells under environmental stress conditions including nutrient-limitation, hyperosmosis, and low temperature, for the storage of metabolic energy and carbon, and also for the consumption of excess energy (e.g., Hirai ...
    Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER
    使用B-PER简单有效地透化蓝藻以进行体内酶分析
    作者:Simon Matthé Erstad and Yumiko Sakuragi日期:01/05/2018,浏览量:172,Q&A: 0
    [Abstract] Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding ...
    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:485,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 ...