Biochemistry

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    Assessing Self-interaction of Mammalian Nuclear Proteins by Co-immunoprecipitation
    Authors:  Claudia Cattoglio, Iryna Pustova, Xavier Darzacq, Robert Tjian and Anders S. Hansen, date: 02/20/2020, view: 3575, Q&A: 0
    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 ...
    Separation and Visualization of Low Abundant Ubiquitylated Forms
    Authors:  Ramona Schuster, Tânia Simões, Fabian den Brave and Mafalda Escobar-Henriques, date: 11/20/2018, view: 3056, Q&A: 0
    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 ...
    Analysis of Direct Interaction between Viral DNA-binding Proteins by Protein Pull-down Co-immunoprecipitation Assay
    Authors:  Ana Lechuga, Mónica Berjón-Otero, Margarita Salas and Modesto Redrejo-Rodríguez, date: 01/05/2018, view: 7016, 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 ...
    Assessment of Modulation of Protein Stability Using Pulse-chase Method
    Author:  Mohamed Elgendy, date: 08/20/2017, view: 8629, Q&A: 0
    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 ...
    Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
    Authors:  Fang Xu, Charles Copeland and Xin Li, date: 07/05/2015, view: 17540, Q&A: 0
    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 ...
    RNA-binding Protein Immunoprecipitation (RIP) to Examine AUF1 Binding to Senescence-Associated Secretory Phenotype (SASP) Factor mRNA
    Authors:  Elise Alspach and Sheila A. Stewart, date: 05/20/2015, view: 12041, Q&A: 0
    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 ...
    Immunoprecipitation of Proteins in Caenorhabditis elegans
    Authors:  Kevin K. Chan, Ashwin Seetharaman, Guillermo Selman and Peter John Roy, date: 04/05/2015, view: 15024, Q&A: 1
    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 ...
    Individual-nucleotide-resolution UV Cross-linking and Immunoprecipitation (iCLIP) of UPF1
    Authors:  David Zünd and Oliver Mühlemann, date: 04/05/2014, view: 12320, Q&A: 0
    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 ...
    UPF1 RNA Immunoprecipitation from Mini-μ Construct–expressing Cells
    Authors:  David Zünd and Oliver Mühlemann, date: 04/05/2014, view: 8859, Q&A: 0
    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 ...
    Co-immunoprecipitation of Flag-TLR3 or Myc-MSR1 with HCV RNA
    Authors:  Daisuke Yamane, Hiromichi Dansako and Stanley M. Lemon, date: 03/05/2014, view: 8032, Q&A: 0
    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 ...



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