Titlebook: Dendritic Cell Protocols; Elodie Segura,Nobuyuki Onai Book 2016Latest edition Springer Science+Business Media New York 2016 culture human

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staging

Isolation and Identification of Conventional Dendritic Cell Subsets from the Intestine of Mice and Mear phagocytes such as macrophages (MFs). Here we detail the protocols we have developed to isolate live leukocytes from both murine and human small and large intestines and describe reliable strategies which can be used to identify bona fide cDCs in such preparations.

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Isolation of Mouse Dendritic Cell Subsets and Macrophages from the Skinlogy upon infection. Here, we describe the procedures to extract those cells from the skin and to analyze them by flow cytometry using a combination of appropriate surface markers allowing further transcriptomic analysis and functional assays.

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Measurement of Export to the Cytosol in Dendritic Cells Using a Cytofluorimetry-Based Assaytive cytosolic substrate of β-lactamase, CCF4. Following uptake of β-lactamase by the DCs, the enzyme undergoes export to the cytosol leading to cleavage of the FRET dye. This cleavage and switch of fluorescence are analyzed by flow cytometry, allowing a quantitative measurement of this event.

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Heinrich Holland,Nandhini Ramanathanmarkers..DC studies might contribute to the comprehension of human immune processes in physiological and pathological conditions. Human blood DCs targeting might be a useful tool to ameliorate inflammatory diseases and improve vaccination strategies.

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