生物化学

分类

    现刊
    Assessing Self-interaction of Mammalian Nuclear Proteins by Co-immunoprecipitation
    共免疫沉淀法评价哺乳动物核蛋白的自身相互作用
    作者:Claudia Cattoglio, Iryna Pustova, Xavier Darzacq, Robert Tjian and Anders S. Hansen日期:02/20/2020,浏览量:890,Q&A: 0
    [Abstract] Protein-protein interactions constitute the molecular foundations of virtually all biological processes. Co-immunoprecipitation (CoIP) experiments are probably the most widely used method to probe both heterotypic and homotypic protein-protein interactions. Recent advances in super-resolution microscopy have revealed that several nuclear proteins ...
    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:1991,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 ...
    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:5345,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 ...
    Assessment of Modulation of Protein Stability Using Pulse-chase Method
    使用脉冲追踪法评估蛋白质稳定性的调节
    作者:Mohamed Elgendy日期:08/20/2017,浏览量:6923,Q&A: 0
    [Abstract] Pulse-chase technique is a method widely used to assess protein or mRNA stability. The principle of pulse-chase relies on labeling proteins or mRNA produced during a short period of time called ‘pulse’ and then following the rate of disappearance of those labeled proteins over a period of time called ‘chase’. This technique thus allows ...
    Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
    采用本氏烟草瞬时表达系统进行蛋白质免疫共沉淀实验
    作者:Fang Xu, Charles Copeland and Xin Li日期:07/05/2015,浏览量:15077,Q&A: 0
    [Abstract] Nicotiana benthamiana (N. benthamiana) is a useful model system to transiently express protein at high level. This protocol describes in detail how to transiently express protein in N. benthamiana and how to carry out protein immunoprecipitation in this expression system. This protocol can be broadly used for ...
    RNA-binding Protein Immunoprecipitation (RIP) to Examine AUF1 Binding to Senescence-Associated Secretory Phenotype (SASP) Factor mRNA
    RNA结合蛋白免疫共沉淀法(RIP)检验结合到衰老相关分泌表型(SASP)因子 的mRNAAUF1
    作者:Elise Alspach and Sheila A. Stewart日期:05/20/2015,浏览量:10664,Q&A: 0
    [Abstract] Immunoprecipitation and subsequent isolation of nucleic acids allows for the investigation of protein:nucleic acid interactions. RNA-binding protein immunoprecipitation (RIP) is used for the analysis of protein interactions with mRNA. Combining RIP with quantitative real-time PCR (qRT-PCR) further enhances the RIP technique by allowing for the ...
    Immunoprecipitation of Proteins in Caenorhabditis elegans
    免疫沉淀法检测秀丽隐杆线虫蛋白质
    作者:Kevin K. Chan, Ashwin Seetharaman, Guillermo Selman and Peter John Roy日期:04/05/2015,浏览量:13073,Q&A: 1
    [Abstract] Immunoprecipitation (IP) is a biochemical technique to precipitate a protein out of solution using an antigen that can specifically bind to that protein. IP can be performed to isolate and concentrate one particular protein from a sample of thousands of different proteins. IP is also readily performed to pull down interacting proteins of complexes ...
    Individual-nucleotide-resolution UV Cross-linking and Immunoprecipitation (iCLIP) of UPF1
    UPF1蛋白单核苷酸分离紫外线交联和免疫沉淀(iCLIP)
    作者:David Zünd and Oliver Mühlemann日期:04/05/2014,浏览量:10975,Q&A: 0
    [Abstract] The fate of mRNA, in particular its stability, localization and rate of translation is regulated by RNA binding proteins assembling to messenger ribonucleoprotein (mRNP) complexes. To investigate the transcriptome-wide RNA binding sites of UPF1, the core factor of nonsense-mediated mRNA decay (NMD), we performed individual-nucleotide-resolution UV ...
    UPF1 RNA Immunoprecipitation from Mini-μ Construct–expressing Cells
    Mini-μ 载体-表达细胞中的UPF1 RNA 免疫沉淀
    作者:David Zünd and Oliver Mühlemann日期:04/05/2014,浏览量:7631,Q&A: 0
    [Abstract] UPF1, an RNA helicase and a core factor of nonsense-mediated mRNA decay (NMD), interacts with RNA independently of the sequence context. To investigate the influence of translation on the association of UPF1 with specific reporter transcripts, UPF1 RNA immunoprecipitations (RIPs) are performed from Hela cells that either express a normally ...
    Co-immunoprecipitation of Flag-TLR3 or Myc-MSR1 with HCV RNA
    Flag-TLR3 或 Myc-MSR1 与 HCV RNA的免疫共沉淀
    作者:Daisuke Yamane, Hiromichi Dansako and Stanley M. Lemon日期:03/05/2014,浏览量:6988,Q&A: 0
    [Abstract] Co-immunoprecipitation assay of TLR3-Flag or Myc-MSR1 with HCV RNA is used to identify direct interaction of viral RNA with host proteins that recognize viral RNA to initiate interferon (IFN) signaling, a crucial antiviral response of the host cells. Both Toll-like receptor 3 (TLR3) and class-A scavenger receptor type 1 (MSR1) proteins recognize ...