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Immunology and Infection

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JoVE Journal Immunology and Infection
Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
 

Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays

Article DOI: 10.3791/57222-v • 11:31 min • July 4th, 2018
July 4th, 2018 •
1, 1, 1, 1
1Institute of Pharmacology, Ruprecht-Karls Heidelberg University

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Here, we present a protocol to isolate and cultivate murine peritoneal mast cells. We also describe two protocols for their functional characterization: a fluorescent imaging of intracellular free Ca2+ concentration and a degranulation assay based on colorimetric quantification of the released β-hexosaminidase.

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Isolation Peritoneum-derived Mast Cells Functional Characterization Ca2+-imaging Degranulation Assays Intracellular Signaling Ion Channels Immune Cell Activation Mediator Release Simplicity Simultaneous Analysis Fura-2 Calcium Imaging Euthanized Mouse 70% Ethanol Foam Block Dorsal Side Down Blunt-edge Scissors Abdomen Peritoneal Cavity Ice-cold RPMI Medium 27 Gauge Needle Epididymal Fat Organ Perforation Peritoneal Cells RMPI Medium
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