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Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
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

DOI:
10.3791/57222-v

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11:31 min

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July 04, 2018

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, , ,
1Institute of Pharmacology,Ruprecht-Karls Heidelberg University

Capitoli

  • 00:04Titolo
  • 00:38PMC Isolation and Cultivation by Intraperitoneal Lavage
  • 03:14Fluorometric Intracellular Free Calcium Concentration Measurement in PMCs
  • 06:38Beta-hexosaminidase Release Measurement in PMCs
  • 08:46Results: Beta-hexosaminidase Release Colorimetric Multi-well Measurements Performed with PMCs Isolated from Wild Type Mice
  • 10:14Conclusion

Summary

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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.

Tags

PeritoneumMast CellsCalcium ImagingDegranulationIntracellular SignalingExcitable CellsIon ChannelsImmune Cell ActivationMediator ReleaseFura-2RPMI MediumPeritoneal CavityCell SuspensionIL-3SCFIgE Receptors

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