Cell Biology

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    Protocols in Current Issue
    Immunohistochemistry of Immune Cells and Cells Bound to in vivo Administered Antibodies in Liver, Lung, Pancreas, and Colon of B6/lpr Mice
    Authors:  Kieran Adam and Adam Mor, date: 07/20/2022, view: 1056, Q&A: 0
    [Abstract]

    Employing a novel mouse model of immune related adverse events (irAEs) induced by combination of anti-PD1 and anti-CTLA-4 antibodies, we visualized immune infiltration into the liver, lung, pancreas, and colon. Here, we describe the avidin-biotin conjugate (ABC) method used to stain T cells (CD4 and CD8), B cells (CD19), macrophages (F4/80), and

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    Flow Cytometric Characterization of Macrophages Infected in vitro with Salmonella enterica Serovar Typhimurium Expressing Red Fluorescent Protein
    [Abstract]

    Macrophages are important for host defense against intracellular pathogens like Salmonella and can be differentiated into two major subtypes. M1 macrophages, which are pro-inflammatory and induce antimicrobial immune effector mechanisms, including the expression of inducible nitric oxide synthase (iNOS), and M2 macrophages, which exert

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    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

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    Fixation and Immunostaining of Endogenous Proteins or Post-translational Modifications in Caenorhabditis elegans
    Authors:  Robert O'Hagan and Irini Topalidou, date: 10/05/2021, view: 2016, Q&A: 0
    [Abstract]

    Although the advent of genetically-encoded fluorescent markers, such as the green fluorescent protein (GFP; Chalfie et al., 1994), has enabled convenient visualization of gene expression in vivo, this method is generally not effective for detecting post-translational modifications because they are not translated from DNA sequences.

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    Preparation of Human Chondrocytes for Profiling Using Cytometry by Time-of-flight (cyTOF)
    Authors:  Fiorella Carla Grandi and Nidhi Bhutani, date: 07/20/2021, view: 2403, Q&A: 0
    [Abstract]

    Single-cell technologies have allowed high-resolution profiling of tissues and thus a deeper understanding of tissue homeostasis and disease heterogeneity. Understanding this heterogeneity can be especially important for tailoring treatments in a patient-specific manner. Here, we detail methods for preparing human cartilage tissue for profiling

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    Tracking the Subcellular Localization of Surface Proteins in Staphylococcus aureus by Immunofluorescence Microscopy
    Authors:  Salvatore J. Scaffidi, Mac A. Shebes and Wenqi Yu, date: 05/20/2021, view: 4798, Q&A: 0
    [Abstract]

    Surface proteins of Staphylococcus aureus and other Gram-positive bacteria play essential roles in bacterial colonization and host-microbe interactions. Surface protein precursors containing a YSIRK/GXXS signal peptide are translocated across the septal membrane at mid-cell, anchored to the cell wall peptidoglycan at the cross-wall compartment,

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    Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila
    Authors:  Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 3219, Q&A: 0
    [Abstract]

    Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to knock down a gene for assessing its function. Similar perturbations in gene expression can be

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    Acidified Blue Ink-staining Procedure for the Observation of Fungal Structures Inside Roots of Two Disparate Plant Lineages
    Authors:  Jill Kowal, Elena Arrigoni and Sophie Lane, date: 10/20/2020, view: 2514, Q&A: 0
    [Abstract] Identifying microscopic mycorrhizal fungal structures in roots, i.e., hyphae, vesicles and arbuscules, requires root staining procedures that are often time consuming and involves chemicals known to present health risks from exposure. By modifying established protocols, our root staining method stains roots using a safe ink- and ...
    Double Labeling of PDGFR-β and α-SMA in Swine Models of Acute Kidney Injury to Detect Pericyte-to-Myofibroblast Transdifferentation as Early Marker of Fibrosis
    [Abstract] Growing evidences suggest that peritubular capillaries pericytes are the main source of scar-forming myofibroblasts during chronic kidney disease (CKD), as well as early phases of acute kidney injury (AKI). In a swine model of sepsis and I/R (Ischemia Reperfusion) injury-induced AKI we demonstrated that renal pericytes are able to ...
    Immunofluorescent Staining of Claudin-2 in Cultured Kidney Tubular Cells
    Authors:  Shaista Anwer and Katalin Szaszi, date: 07/20/2020, view: 2741, Q&A: 0
    [Abstract] Members of the claudin family of tight junction proteins regulate paracellular permeability and modulate cell signaling. During junction remodeling, these proteins are selectively inserted into or retrieved from the tight junctions, but the control and coordination of these processes remain incompletely understood. Visualization of claudins allows ...



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