13,524 Result(s)
within
Cell Biology
Yu** **
Protocol
Life Sciences
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Protocol
Affinity Purification of Intraflagellar Transport (IFT) Proteins in Mice Using Endogenous Streptavidin/FLAG Tags
Biological complexity is achieved through elaborate interactions between relatively few individual components. Affinity purification (AP) has allowed these networks of protein-protein interactions that regulat...
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Controlling the Subcellular Localization of Signaling Proteins Using Chemically Induced Dimerization and Optogenetics
A given protein can perform numerous roles in a cell with its participation in protein complexes and distinct localization within the cell playing a critical role in its diverse functions. Thus, the ability to...
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Measurement of Intracellular Ca2+ for Studying GPCR-Mediated Lipid Signaling
Intracellular Ca2+ can be conveniently monitored by sensitive Ca2+ fluorescent dyes in live cells. The Gαq involved lipid signaling pathways and, thus, can be studied by intracellular Ca2+ imaging. Here we descri...
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Atomistic Simulations of Sarcomere Proteins
Concerted atomic motions are requisite for sarcomere protein function and may become disrupted in HCM pathologies. Computational approaches such as molecular dynamics simulation can resolve such dynamics with ...
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Monitoring of Inflammasome Activation of Macrophages and Microglia In Vitro, Part 2: Assessing Inflammasome Activation
are macromolecular complexes that assemble upon the detection of cytoplasmic pathogen-associated or danger-associated signals and induce a necrotic type of cell death termed pyroptosis , facilitating pro-infl...
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Probing Bacterial Cell Division and Cell Envelope Biogenesis with Live-Cell Fluorescence Microscopy
The development of advanced microscopy techniques has ushered in a new era of research as it helps understand biological processes on a deeper, mechanistic, and molecular level like never before. Live-cell flu...
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Automated Cell Counting of Macrophages In Situ
Using the open-source image analysis software CellProfiler to automatically quantify antibody-stained or fluorescently labeled macrophages in situ provides accurate and reproducible cell counts. It is a vastly...
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Isolation and Purification of Viable PGCs from Mouse Embryos
In all organisms with sexual reproduction, sperm and oocytes derive from embryonic precursors termed primordial germ cells (PGCs) which pass on genetic information to subsequent generations. Studies aimed to u...
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Isolation of Human Adipose-Derived Stem Cells
Human adipose-derived stromal/stem cells (hASCs) are a promising source of adult stem cells used in numerous applications in regenerative medicine. We present the protocols from our laboratory for isolating an...
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Direct Detection of Borrelia Species in Tissues
Among the controversies in Lyme disease is the potential for Borrelia spirochetes to persist after guideline-directed antimicrobial therapy. Direct detection of the spirochetes has been essential to explore this ...
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Workflow for Performing Genetic Manipulation in Human Trophoblast Stem Cells Using CRISPR/Cas9 Technology
Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based tool kits greatly facilitate the investigation of gene function, especially gene editing and transcriptional regulation technology....
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MAARS Software for Automatic and Quantitative Analysis of Mitotic Progression
In this chapter, we describe a software called MAARS (Mitotic Analysis And Recording System) that enables automatic and quantitative analysis of mitotic progression on an open-source platform. This computer-as...
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Isolation and Maintenance in Culture of Primary Human Trophoblast from Term Placentae
Trophoblasts are placenta-specific epithelial cells that play an essential role in conducting nutrient, gas, and waste exchange between the fetus and the mother. Primary culture of human trophoblasts from dona...
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SARS-CoV-2 Infection of Human Primary Nasal Multiciliated Epithelial Cells Grown on Air-Liquid Interface Cultures
Respiratory epithelial cells fail to exhibit natural phenotypic and morphological characteristics when grown in standard cell culture conditions. To better understanding respiratory pathogen host-cell interact...
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Liposome-Encapsulated Anti-inflammatory Proteins for Targeted Delivery to the Placenta to Treat Fetal Growth Restriction
Fetal growth restriction (FGR), the failure of a fetus to reach its genetically determined growth potential, is a serious complication affecting up to 10% of pregnancies. FGR is a major risk factor for stillbi...
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3D Breast Cancer Model on Silk Fibroin–Integrated Microfluidic Chips
To imitate in vivo environment of cells, microfluidics offer controllable fashions at micro-scale and enable regulate flow-related parameters precisely, leveraging the current state of 3D systems to 4D level t...
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Investigating Epidermal Growth Factor Receptor Trafficking with High-Resolution Enzymatic Protein-Tagging and Transmission Electron Microscopy
Enzymatic ascorbate peroxidase (APEX) tagging allows for high-resolution, three-dimensional protein distribution analyses in cells and tissues. This chapter describes the application of APEX-tagging to visuali...
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Quantitative Analysis of Whole-Mount Fluorescence-Stained Tumor Spheroids in Phenotypic Drug Screens
Three-dimensional cell cultures, such as spheroids or organoids, serve as important models for drug screening purposes. Optical tissue clearing (OTC) enhances the visualization of fluorescence stainings and en...
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BMI1 Transduction of Human Airway Epithelial Cells for Expansion of Proliferation and Differentiation
Air-liquid interface (ALI)-cultured cells are widely used as in vitro models of the human respiratory airway in studies of pulmonary physiology, disease, and therapies. However, the primary basal cells require...
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Effects of a Laparotomy on Targeted Lipidomics Profiles in a Mouse Model of Surgical Stress During Aging
Laparotomy (EL) is one of the most common procedures performed among surgical specialties. Previous research demonstrates that surgery is associated with an increased inflammatory response. Low psoas muscle ma...
