Measuring in vitro ATPase Activity with High Sensitivity Using Radiolabeled ATP
    Authors:  Sarina Veit and Thomas Günther Pomorski, date: 05/20/2023, view: 203, Q&A: 0

    ATPase assays are a common tool for the characterization of purified ATPases. Here, we describe a radioactive [γ-32P]-ATP-based approach, utilizing complex formation with molybdate for phase separation of the free phosphate from

    In vitro Reconstitution of Phase-separated p62 Bodies on the Arp2/3-derived Actin Network
    Authors:  Tong Liu, Mengbo Xu and Na Mi, date: 04/20/2023, view: 320, Q&A: 0

    In cells, p62/SQSTM1 undergoes liquid–liquid phase separation (LLPS) with poly-ubiquitin chains to form p62 bodies that work as a hub for various cellular events, including selective autophagy. Cytoskeleton components such as Arp2/3-derived

    Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)

    Lipid droplets (LD), triglycerides and sterol esters among them, are well known for their capacity as lipid storage organelles. Recently, they have emerged as critical cytoplasmic structures involved in numerous biological functions. LD storage is

    Imaging Membrane Proteins Using Total Internal Reflection Fluorescence Microscopy (TIRFM) in Mammalian Cells
    Authors:  Kirin D. Gada, Jordie M. Kamuene, Takeharu Kawano and Leigh D. Plant, date: 02/20/2023, view: 481, Q&A: 0

    The cell surfaceome is of vital importance across physiology, developmental biology, and disease states alike. The precise identification of proteins and their regulatory mechanisms at the cell membrane has been challenging and is typically

    Lysate-to-grid: Rapid Isolation of Native Complexes from Budding Yeast for Cryo-EM Imaging

    Single-particle electron cryo-microscopy (cryo-EM) is an effective tool to determine high-resolution structures of macromolecular complexes. Its lower requirements for sample concentration and purity make it an accessible method to determine

    Focused Ion Beam Milling and Cryo-electron Tomography Methods to Study the Structure of the Primary Cell Wall in Allium cepa
    Authors:  William J. Nicolas, Grant J. Jensen and Elliot M. Meyerowitz, date: 12/05/2022, view: 627, Q&A: 0

    Cryo-electron tomography (cryo-ET) is a formidable technique to observe the inner workings of vitrified cells at a nanometric resolution in near-native conditions and in three-dimensions. One consequent drawback of this technique is the sample

    A Fast and Efficient Decellularization Method for Tissue Slices

    The study and use of decellularized extracellular matrix (dECM) in tissue engineering, regenerative medicine, and pathophysiology have become more prevalent in recent years. To obtain dECM, numerous decellularization procedures have been developed

    In situ cryo-FIB/SEM Specimen Preparation Using the Waffle Method

    Cryo-focused ion beam (FIB) milling of vitrified specimens is emerging as a powerful method for in situ specimen preparation. It allows for the preservation of native and near-native conditions in cells, and can reveal the molecular structure of

    Binding Affinity Measurements Between DNA Aptamers and their Virus Targets Using ELONA and MST
    Authors:  Gregory T. Pawel, Yuan Ma, Yuting Wu, Yi Lu and Ana Sol Peinetti, date: 11/05/2022, view: 807, Q&A: 0

    Aptamers have been selected with strong affinity and high selectivity for a wide range of targets, as recently highlighted by the development of aptamer-based sensors that can differentiate infectious from non-infectious viruses, including human

    A Fluorescence-based Approach Utilizing Self-labeling Enzyme Tags to Determine Protein Orientation in Large Unilamellar Vesicles
    Authors:  Laura Charlotte Paweletz, Sarina Veit and Thomas Günther Pomorski, date: 11/05/2022, view: 640, Q&A: 0

    Reconstitution of membrane proteins into large unilamellar vesicles is an essential approach for their functional analysis under chemically defined conditions. The orientation of the protein in the liposomal membrane after reconstitution depends on


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