Protocols in Current Issue
    Immunohistochemical Staining of CD8α in Diabetic Mouse Kidney
    Authors:  Ming-Zhi Zhang and Raymond C. Harris, date: 09/20/2019, view: 911, 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 ...
    Immunohistochemical Identification of Human Skeletal Muscle Macrophages
    [Abstract] Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage ...
    Thermal Stability of Heterotrimeric pMHC Proteins as Determined by Circular Dichroism Spectroscopy
    [Abstract] T cell receptor (TCR) recognition of foreign peptide fragments, presented by peptide major histocompatibility complex (pMHC), governs T-cell mediated protection against pathogens and cancer. Many factors govern T-cell sensitivity, including the affinity of the TCR-pMHC interaction and the stability of pMHC on the surface of antigen presenting ...
    Lung Section Staining and Microscopy
    Authors:  Xiaofeng Zhou and Bethany B Moore, date: 05/20/2017, view: 15276, Q&A: 0
    [Abstract] Our protocol describes immunofluorescent staining, hematoxylin and eosin staining and Masson’s trichrome staining on lung sections.
    Cell Tracer Violet and CellTracker Red CMTPX Staining of Purified Mature Plasmodium-infected Red Blood Cells
    [Abstract] Efficient staining methods to identify Plasmodium-infected red blood cells (iRBCs) are crucial to discriminate precisely the immune cells responsible for their elimination from circulation. Here, we describe the protocol for the purification of iRBCs and their subsequent staining with the vital dyes Cell Tracer Violet (CTV) or CellTracker ...
    Quantification of Rae-1 Staining Intensity in Glomeruli
    Authors:  Roberto Spada and Domingo F. Barber, date: 03/05/2016, view: 4887, Q&A: 0
    [Abstract] NKG2D is expressed on all NK cells and on subsets of NKT, CD8, CD4 and γδ T cells. NKG2D is activated by NKG2D ligands, a stress-induced family of MHC-I-like proteins. These ligands are upregulated on stressed/infected cells and are not widely expressed on healthy adult tissue. NKG2D ligands have been widely studied as potential targets for ...
    Cytokine-stimulated Phosphoflow of PBMC Using CyTOF Mass Cytometry
    Authors:  Rosemary Fernandez and Holden Maecker, date: 06/05/2015, view: 12671, Q&A: 1
    [Abstract] Phosphorylation of tyrosine, serine, and threonine residues is critical for the control of protein activity involved in various cellular events. An assortment of kinases and phosphatases regulate intracellular protein phosphorylation in many different cell signaling pathways. These pathways include T and B cell signaling, regulating growth and ...
    Cytokine-Stimulated Phosphoflow of Whole Blood Using CyTOF Mass Cytometry
    Authors:  Rosemary Fernandez and Holden Maecker, date: 06/05/2015, view: 9987, Q&A: 1
    [Abstract] The ability to assess the function of a range of cytokine, antigen receptor, and Toll-like receptor (TLR) signaling pathways in a range of immune cells could provide a kind of fingerprint of the state of the human immune system. The mass cytometry or CyTOF, platform allows for the parallel application of about 40 labeled antibodies to a single ...
    Phenotyping of Live Human PBMC using CyTOFTM Mass Cytometry
    Authors:  Michael D. Leipold and Holden Maecker, date: 01/20/2015, view: 13143, Q&A: 3
    [Abstract] Single-cell analysis has become an method of importance in immunology. Fluorescence flow cytometry has been a major player. However, due to issues such as autofluorescence and emission spillover between different fluorophores, alternative techniques are being developed. In recent years, mass cytometry has emerged, wherein antibodies labeled with ...
    Immunocytochemical Detection of Recombinant Biomphalysin on Schistosoma mansoni Sporocysts
    Authors:  David Duval, Richard Galinier, Julien Portela, Guillaume Mitta and Benjamin Gourbal, date: 11/20/2013, view: 6797, Q&A: 0
    [Abstract] Schistosomiasis, or bilharzia, is a tropical disease caused by worms of the genus Schistosoma which infect about 200 million people. The life cycle of the parasite requires Biomphalaria, a specific genus of freshwater snails, as intermediate. Using an interactome approach employing B. glabrata plasma and S. mansoni ...

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