Protocols in Current Issue
    Analysis of Generalized Fibrosis in Mouse Tissue Sections with Masson’s Trichrome Staining
    Authors:  Diantha van de Vlekkert, Eda Machado and Alessandra d’Azzo, date: 05/20/2020, view: 516, Q&A: 0
    [Abstract] Expansion of fibrous connective tissue and abnormal deposition of extracellular matrix (ECM) are at the basis of many fibrotic diseases. Fibrosis can occur in response to both physiological and pathological cues, including wound healing, tissue remodeling/repair and inflammation. Chronic fibrosis can lead to severe tissue damage, organ failure and ...
    Measuring Small-molecule Inhibition of Protein Interactions in Live Cells Using FLIM-FRET
    Authors:  James M. Pemberton, Qian Liu and David W. Andrews, date: 10/20/2019, view: 2376, Q&A: 0
    [Abstract] This protocol was designed to quantitatively measure small-molecule displacement of proteins in live mammalian cells using fluorescence lifetime imaging microscopy–Förster resonance energy transfer (FLIM-FRET). Tumour cell survival is often dependent on anti-apoptotic proteins, which bind to and inhibit pro-apoptotic proteins, thus preventing ...
    Isolation of Pure Mitochondria from Rat Kidneys and Western Blot of Mitochondrial Respiratory Chain Complexes
    [Abstract] Cardiac, neuronal and renal tubular epithelial cells are the most metabolically active cells in the body. Their fate depends largely on their mitochondria as the primary energy generating system which participates in the control of apoptosis, cell cycle and metabolism. Thus, mitochondrial dysfunction is a hallmark of many chronic diseases ...
    Immunohistochemical Staining of CD8α in Diabetic Mouse Kidney
    Authors:  Ming-Zhi Zhang and Raymond C. Harris, date: 09/20/2019, view: 679, Q&A: 0
    [Abstract] Immune cell infiltration, particularly cytotoxic CD8α lymphocyte infiltration, plays an important role in development of diabetic nephropathy. Although CD8α infiltration can be evaluated by its production of cytokines, its localization in the kidney is of particular importance. The current protocol describes CD8α immunostaining using a Vectastain ...
    Immunofluorescence Staining of WT-1/Podocalyxin on Mouse Kidney Sections
    Authors:  Carole Henique, Olivia Lenoir and Pierre-Louis Tharaux, date: 04/20/2019, view: 1848, Q&A: 0
    [Abstract] In glomerular diseases, podocytes are one type of cells involved in dysfunction of glomerular filtration. In these conditions, podocyte proteins expression change. Therefore, immunofluorescent staining of podocytes can be performed to visualize podocyte localization of proteins of interest. In this protocol, we detail a method to stain podocytes ...
    Quantifying Podocytes and Parietal Epithelial Cells in Human Urine Using Liquid-based Cytology and WT1 Immunoenzyme Staining
    [Abstract] In glomerular disease, podocytes and parietal epithelial cells (PECs) are shed in the urine. Therefore, urinary podocytes and PECs are noninvasive biomarkers of glomerular disease. The purpose of this protocol is to employ immunocytochemistry to detect podocytes and PECs, using the WT1 antibody on liquid-based cytology slides.
    Murine Pancreatic Islets Transplantation under the Kidney Capsule
    [Abstract] Type 1 diabetes (T1D) is an autoimmune disease caused by the lack of insulin-producing pancreatic beta cells leading to systemic hyperglycemia. Pancreatic islet transplantation is a valid therapeutic approach to restore insulin loss and to promote adequate glycemic control. Pancreatic islet transplantation in mice is an optimal preclinical model ...
    Sebinger Culture: A System Optimized for Morphological Maturation and Imaging of Cultured Mouse Metanephric Primordia
    Authors:  Mona Elhendawi and Jamie A. Davies, date: 02/20/2018, view: 3091, Q&A: 0
    [Abstract] Here, we present a detailed protocol on setting up embryonic renal organ cultures using a culture method that we have optimised for anatomical maturation and imaging. Our culture method places kidney rudiments on glass in a thin film of medium, which results in very flat cultures with all tubules in the same image plane. For reasons not yet ...
    Analysis of the Virulence of Uropathogenic Escherichia coli Strain CFT073 in the Murine Urinary Tract
    Authors:  Anna Waldhuber, Manoj Puthia, Andreas Wieser, Catharina Svanborg and Thomas Miethke , date: 02/05/2017, view: 6865, Q&A: 0
    [Abstract] This urinary tract infection model was used to monitor the efficacy of a new virulence factor of the uropathogenic Escherichia coli strain CFT073 in vivo. The new virulence factor which we designated TIR-containing protein C (TcpC) blocks Toll-like receptor signaling and the NLRP3 inflammasome signaling cascade by interacting ...
    Process and Analysis of Kidney Infiltrates by Flow Cytometry from Murine Lupus Nephritis
    Authors:  Ramalingam Bethunaickan and Anne Davidson, date: 05/05/2012, view: 14536, Q&A: 0
    [Abstract] Methods for the isolation and characterization of mononuclear phagocytes from the kidneys of mice with SLE are essential to understand the patho-physiology of the disease. Activation of these cells is associated with the onset of clinical disease in mice and infiltration with these cells is associated with poor prognosis in humans.An analysis of ...

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