Ultrasensitive Detection of Attomolar Protein Concentrations by Dropcast Single Molecule Assays [PDF]
Journal of the American Chemical Society, 2020 Measurements of very low levels of biomolecules, including proteins and nucleic acids, remain a critical challenge in many clinical diagnostic applications due to insufficient sensitivity. While digital measurement methods such as Single Molecule Arrays (Simoa), or digital ELISA, have made significant advances in sensitivity, there are still many ...
Padric M Garden, David Walt, Connie Wu
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Direct single-molecule imaging for diagnostic and blood screening assays [PDF]
Proceedings of the National Academy of Sciences of the United States of America, 2021 Significance Direct imaging and counting single molecules captured on magnetic microparticles enables diagnostic assays with ultrahigh sensitivities and wide linear detection ranges. The method offers simple assay protocols, short incubation times, minimal reagent consumption, and does not require enzymatic signal amplification.
Patrick Macdonald +2 more
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Cytoplasmic Actin: Purification and Single Molecule Assembly Assays [PDF]
Methods in Molecular Biology, 2013 The actin cytoskeleton is essential to all eukaryotic cells. In addition to playing important structural roles, assembly of actin into filaments powers diverse cellular processes, including cell motility, cytokinesis, and endocytosis. Actin polymerization is tightly regulated by its numerous cofactors, which control spatial and temporal assembly of ...
Scott Hansen +2 more
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Single molecule enzyme-linked immunosorbent assays: Theoretical considerations [PDF]
Journal of Immunological Methods, 2012 We have developed a highly sensitive immunoassay-called digital ELISA-that is based on the detection of single enzyme-linked immunocomplexes on beads that are sealed in arrays of femtoliter wells. Digital ELISA was designed to be highly efficient in the capturing of target proteins, labeling of these proteins, and their detection in single molecule ...
David Duffy +2 more
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DNA Flow-Stretch Assays for Studies of Protein-DNA Interactions at the Single-Molecule Level
Applied Nano, 2022 Protein-DNA interactions are the core of the cell’s molecular machinery. For a long time, conventional biochemical methods served as a powerful investigatory basis of protein-DNA interactions and target search mechanisms.
Aurimas Kopūstas +2 more
doaj +3 more sources
Protocol for measuring endocrine-disruptive effects on transcriptional bursting using single-molecule imaging in human breast cancer cells [PDF]
STAR ProtocolsSummary: Transcriptional bursts regulate gene expression by altering burst size or burst frequency. Here, we present a protocol to analyze estrogen-responsive transcriptional bursting of the TFF1 gene in human breast cancer cell lines by integrating ...
Pelin Yaşar +2 more
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High-Resolution Single-Molecule Kinesin Assays at kHz Frame Rates
Methods in Molecular Biology, 2018 This chapter describes methods for high-speed, unloaded, in vitro single-molecule kinesin tracking experiments. Instructions are presented for constructing a total internal reflection dark-field microscope (TIRDFM) and labeling motors with gold nanoparticles. An AMP-PNP unlocking assay is introduced as a specialized means of capturing processive events
William Hancock, Keith Mickolajczyk
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Protocol for single-molecule pull-down of fluorescently tagged oligomers from cell lysates
STAR Protocols, 2023 Summary: Mutations in intrinsically disordered proteins drive the irreversible formation of pathological aggregates, a hallmark of neurodegenerative diseases.
Nathalie Djaja, Sua Myong
doaj +1 more source
Quantifying transport dynamics with three-dimensional single-particle tracking in adherent cells
STAR Protocols, 2022 Summary: Intracellular transport plays an important role in maintaining the physiological functions of cells. Here, we describe a protocol for 3D single-particle tracking within living cells.
Chao Jiang +3 more
doaj +1 more source
Protocol for generation and regeneration of PEG-passivated slides for single-molecule measurements
STAR Protocols, 2022 Summary: Single-molecule fluorescence detection by total internal reflection microscope requires surface passivation by polyethylene glycol (PEG) coating, which is labor intensive and is only good for one or two experiments. Here, we present an efficient
Tapas Paul, Sua Myong
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