Protocols in Current Issue
    Optogenetic Tuning of Protein-protein Binding in Bilayers Using LOVTRAP
    Authors:  Doug Tischer and Orion D. Weiner, date: 09/05/2020, view: 650, Q&A: 0
    [Abstract] Modern microscopy methods are powerful tools for studying live cell signaling and biochemical reactions, enabling us to observe when and where these reactions take place from the level of a cell down to single molecules. With microscopy, each cell or molecule can be observed both before and after a given perturbation, facilitating better inference ...
    Contemporaneous Measurement of Outer and Inner Membrane Permeability in Gram-negative Bacteria
    Authors:  Bo Ma, Chao Fang, Jing Zhang, Mingzhi Wang, Xiaoxing Luo and Zheng Hou, date: 03/05/2020, view: 1088, Q&A: 0
    [Abstract] The emergence and rapid spread of multidrug resistance in bacteria have led to the urgent need for novel antibacterial agents. Membrane permeabilization is the mechanism for many antibacterial molecules that are being developed against gram-negative bacteria. Thus, to determine the efficacy of a potential antibacterial molecule, it is important to ...
    Photoactivable Cholesterol as a Tool to Study Interaction of Influenza Virus Hemagglutinin with Cholesterol
    Authors:  Bodan Hu, Mohamed Rasheed Gadalla, Christoph Thiele and Michael Veit, date: 02/20/2020, view: 1348, Q&A: 0
    [Abstract] Non-covalent binding of cholesterol to the transmembrane region of proteins affect their functionalities, but methods to prove such an interaction are rare. We describe our protocol to label the hemagglutinin (HA) of Influenza virus with a cholesterol derivative in living cells or with immunoprecipitated protein. We synthesized a “clickable” ...
    Protein-lipid Interaction Analysis by Surface Plasmon Resonance (SPR)
    Authors:  Olga Lucia Baron and David Pauron, date: 09/20/2014, view: 9798, Q&A: 0
    [Abstract] Interactions of lipids with proteins are essential events in the framework of biological membranes. Assessment of the affinity and specificity of protein-lipid binding can give useful information to elucidate cell membrane functions. Surface Plasmon Resonance (SPR) is a powerful technology to study macromolecular interactions, allowing direct and ...
    Quantitative Analysis of Cellular Diacylglycerol Content
    Authors:  Fikadu G. Tafesse, Karin Strijbis and Hidde L. Ploegh, date: 08/05/2014, view: 9140, Q&A: 0
    [Abstract] Diacylglycerol (DAG) is a bioactive lipid with diverse biological roles. DAG transiently accumulates in a membrane upon receipt of an appropriate stimulus that activates phospholipase C to cleave phospholipids. The resulting hydrolysis product DAG binds to proteins such as protein kinase C to initiate a variety of downstream cellular processes. ...
    Dye Release Experiments with Dextran Loaded Vesicles
    Authors:  Marc-Antoine Sani, Neil M. O’Brien-Simpson and Frances Separovic, date: 07/20/2014, view: 7369, Q&A: 0
    [Abstract] Dye release experiments are a widely used method to assess the interactions between membrane-active molecules and lipid membranes. Of particular interest is the ability to assess the degree of the lipid bilayer perturbation by simultaneously encapsulating dye of different sizes, such as dextrans grafted with a chromophore. In this assay, dextran ...
    Phosphoinositides Coated Beads Binding Assay
    [Abstract] The PIs coated beads assay or “PIP-Beads” developed by Echelon Biosciences (Salt Lake City, USA) is a quick assay to recognize which PIs are able to bind to a given protein domain, in a quantitative way. It is much faster and cheaper than liposomes and more reproducible than PIP-strip assays. The “PIP-Beads” assay is a biochemical assay that ...
    PTEN-lipid Binding Assay
    Authors:  Sridhar Kavela, Swapnil R. Shinde and Subbareddy Maddika, date: 08/20/2013, view: 7820, Q&A: 0
    [Abstract] The lipid and protein interactions are an integral and important part of many cellular signaling pathways. The understanding of the selective and specific interaction of the given lipid molecule with the target protein is required for studying cellular signaling. In this assay, different lipids are spotted onto a nitrocellulose membrane to which ...

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