Biochemistry

Categories

    Protocols in Current Issue
    Immunoisolation of Endosomal Recycling Vesicles from Saccharomyces cerevisiae
    Authors:  Sho W. Suzuki and Scott D. Emr, date: 05/05/2022, view: 620, Q&A: 0
    [Abstract]

    Endosomal recycling is essential for the appropriate function of the endosome. During this process, endosomal coat complexes (i.e., retromer, and Mvp1) are recruited to the endosome, and deform its membrane to form recycling vesicles. To further analyze this, we developed a protocol for the immunoisolation of recycling vesicles from budding yeast.

    ...
    Analysis of Heterocyst and Akinete Specific Glycolipids in Cyanobacteria Using Thin-layer Chromatography
    Authors:  Ritu Garg, Rebeca Perez and Iris Maldener, date: 03/20/2022, view: 658, Q&A: 0
    [Abstract]

    Several filamentous cyanobacteria like Nostoc differentiate specialized cells in response to changes in environmental factors, such as low light or nutrient starvation. These specialized cells are termed heterocysts and akinetes. Under conditions of nitrogen limitation, nitrogen-fixing heterocysts form in a semi-regular pattern and provide the

    ...
    A Fluorescence-based Assay for Measuring Phospholipid Scramblase Activity in Giant Unilamellar Vesicles
    Authors:  Patricia P. M. Mathiassen and Thomas Günther Pomorski, date: 03/20/2022, view: 712, Q&A: 0
    [Abstract]

    Transbilayer movement of phospholipids in biological membranes is mediated by a diverse set of lipid transporters. Among them are scramblases that facilitate rapid bi-directional movement of lipids without metabolic energy input. In this protocol, we describe the incorporation of phospholipid scramblases into giant unilamellar vesicles (GUVs)

    ...
    Rapid Lipid Quantification in Caenorhabditis elegans by Oil Red O and Nile Red Staining
    [Abstract]

    The ability to stain lipid stores in vivo allows for the facile assessment of metabolic status in individuals of a population following genetic and environmental manipulation or pharmacological treatment. In the animal model Caenorhabditis elegans, lipids are stored in and mobilized from intracellular lipid droplets in the intestinal and

    ...
    Reconstitution of Membrane-associated Components of a G-protein Signaling Pathway on Membrane-coated Nanoparticles (Lipobeads)
    Authors:  Michael J. Irwin, Xin Wang and Rick H. Cote, date: 01/20/2022, view: 866, Q&A: 0
    [Abstract]

    G-protein coupled signaling pathways are organized into multi-protein complexes called signalosomes that are located within and on cellular membranes. We describe the use of silica nanoparticles coated with a unilamellar phospholipid bilayer (lipobeads) to reconstitute the activated photoreceptor G-protein α-subunit (Gtα*) with its

    ...
    A Simple and Straightforward Approach for Generating Small, Stable, Homogeneous, Unilamellar 1-Palmitoyl 2-Oleoyl Phosphatidylcholine (POPC) Bilayer Vesicles
    Authors:  Yong-Guang Gao, Le Thi My Le, Amer Alam and Rhoderick E. Brown, date: 12/20/2021, view: 1428, Q&A: 0
    [Abstract]

    Various methods have been developed to generate phosphoglyceride liposomes. Approaches resulting in homogeneous populations of unilamellar bilayer vesicles are generally preferred to mimic various cell membrane situations, as well as to optimize aqueous solute trapping efficiency using the least amount of lipid for biotechnological purposes. Most

    ...
    Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts
    [Abstract]

    The efficient ATP production in mitochondria relies on the highly specific organization of its double membrane. Notably, the inner mitochondrial membrane (IMM) displays a massive surface extension through its folding into cristae, along which concentrate respiratory complexes and oligomers of the ATP synthase. Evidence has accumulated to highlight

    ...
    Kinetic Analysis of a Protein-protein Complex to Determine its Dissociation Constant (KD) and the Effective Concentration (EC50) of an Interplaying Effector Molecule Using Bio-layer Interferometry
    Authors:  Tim Orthwein, Luciano F. Huergo, Karl Forchhammer and Khaled A. Selim, date: 09/05/2021, view: 2455, Q&A: 0
    [Abstract]

    Biolayer interferometry (BLI) is an emerging analytical tool that allows the study of protein complexes in real time to determine protein complex kinetic parameters. This article describes a protocol to determine the KD of a protein complex using a 6×His tagged fusion protein as bait immobilized on the NTA sensor chip of the

    ...
    Oil Red O Staining for Lipid Content in Caenorhabditis elegans
    Authors:  Feng-Yung Wang and Tsui-Ting Ching, date: 08/20/2021, view: 2534, Q&A: 0
    [Abstract]

    The nematode Caenorhabditis elegans has emerged as a popular model system for studying the regulation of lipid metabolism. Therefore, it is critical to develop a method for determining fat storage in individual worms. Oil Red O (ORO) staining has been validated as an accurate assessment for major fat storage in C. elegans. Here, we describe an

    ...
    Yeast Lipid Extraction and Analysis by HPTLC
    Authors:  Dan Li, Zheng-Tan Zhang, Cheng-Wen He and Zhiping Xie, date: 07/05/2021, view: 2211, Q&A: 0
    [Abstract]

    The diversity of lipid structures, properties, and combinations in biological tissues makes their extraction and analysis an experimental challenge. Accordingly, even for one of the simplest single-cellular fungi, the budding yeast (Saccharomyces cerevisiae), numerous extraction and analysis protocols have been developed to separate and quantitate

    ...



    We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.