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    Protocols in Current Issue
    Mating Based Split-ubiquitin Assay for Detection of Protein Interactions
    Authors:  Wijitra Horaruang and Ben Zhang, date: 05/05/2017, view: 6217, Q&A: 0
    [Abstract] The mating based split-ubiquitin (mbSUS) assay is an alternative method to the classical yeast two-hybrid system with a number of advantages. The mbSUS assay relies on the ubiquitin-degradation pathway as a sensor for protein-protein interactions, and it is suitable for the determination of interactions between full-length proteins that are ...
    Protein Immunoprecipitation Using Nicotiana benthamiana Transient Expression System
    Authors:  Fang Xu, Charles Copeland and Xin Li, date: 07/05/2015, view: 11824, 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 ...
    A Chemiluminescence Based Receptor-ligand Binding Assay Using Peptide Ligands with an Acridinium Ester Label
    Authors:  Mari Wildhagen, Melinka A. Butenko, Reidunn B. Aalen, Georg Felix and Markus Albert, date: 03/20/2015, view: 6640, Q&A: 0
    [Abstract] Studying the biochemical interaction of ligands with their corresponding receptors requires highly sensitive detection and monitoring of the bound ligand. Classically, radioactively labelled ligands have been widely used as highly sensitive tools for such binding measurements. Disadvantages of radiolabelling include instability of products, high ...
    Phosphatase Protection Assay: 14-3-3 Binding Protects the Phosphate group of RSG from λ Protein Phosphatase
    Authors:  Takeshi Ito and Yohsuke Takahashi, date: 02/05/2015, view: 6109, Q&A: 0
    [Abstract] 14-3-3 proteins regulate diverse cellular processes in eukaryotes by binding to phospho-serine or threonine of target proteins. One of the physiological functions of 14-3-3 is to bind and protect phosphate groups of the target proteins against phosphatases. REPRESSION OF SHOOT GROWTH (RSG) is a tobacco (Nicotiana tabacum) transcription ...
    CytoTrap Two-Hybrid Screening Assay
    Authors:  Lei Zhang, Liqun Du and B. W. Poovaiah, date: 12/05/2014, view: 7796, Q&A: 0
    [Abstract] CytoTrap two-hybrid system provides an alternate strategy to detect protein-protein interactions in yeast. In this system, bait protein is fused with human son of sevenless (hSos) protein (Li et al., 1993), and a cDNA library or prey protein is expressed by fusion with myristoylation signal which anchors the prey fusion protein to yeast ...
    SGR-based Reporter to Assay Plant Transcription Factor-promoter Interactions
    Authors:  Shisong Ma and Savithramma Dinesh-Kumar, date: 08/20/2014, view: 6543, Q&A: 0
    [Abstract] We developed an in vivo method to assay plant transcription factor (TF)–promoter interactions using the transient expression system in Nicotiana benthamiana (N. benthamiana) plants. The system uses the Arabidopsis stay green (SGR) gene as a reporter. Induction of SGR expression in N. ...
    Binding Assay of Cytosolic Proteins to the Cytoskeleton
    Authors:  Stefano Del Duca and Giampiero Cai, date: 11/20/2013, view: 7527, Q&A: 1
    [Abstract] Many cellular proteins interact with the cytoskeleton (both actin filaments and microtubules), either dynamically or permanently. This interaction is required during different aspects of the cell life, for example during the process of cell division. In addition, many enzymes interact transiently with actin filaments and microtubules in order to ...
    Analysis of RNA-protein Interactions Using Electrophoretic Mobility Shift Assay (Gel Shift Assay)
    Authors:  Saiprasad Goud Palusa and Anireddy S. N. Reddy, date: 11/20/2013, view: 18665, Q&A: 5
    [Abstract] RNA binding proteins (RBPs) play a crucial role in regulating gene expression at the post-transcriptional level at multiple steps including pre-mRNA splicing, polyadenylation, mRNA stability, mRNA localization and translation. RBPs regulate these processes primarily by binding to specific sequence elements in nascent or mature transcripts. There ...
    Northwestern Blot of Protein-RNA Interaction from Young Rice Panicles
    Authors:  Saminathan Thangasamy and Guang-Yuh Jauh, date: 04/05/2013, view: 10138, Q&A: 0
    [Abstract] The northwestern assay is employed to study the interaction between protein and RNA. The RNA binding proteins tend to bind to different kinds of RNA through either known domains or unknown sequences of proteins. Rice LGD1 recombinant protein, a grass-specific novel protein with RNA binding sequences in its C-terminal, was used to probe its ...
    UV Cross-linking Assay and Competition Assay
    Authors:  Ying-Wen Huang and Yau-Heiu Hsu, date: 12/20/2012, view: 12009, Q&A: 1
    [Abstract] UV cross-linking assay is a standard method used to detect protein-RNA interaction. This method takes advantage of UV irradiation to trigger the formation of the covalently bonded RNP (ribonucleoprotein) complex that is more stable and makes it possible to be isolated in the denaturing conditions. Briefly, 32P-labeled RNA probe and ...



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