Abstract
Mast cell activation is a central process in the initiation of allergic disorders. As described elsewhere in this volume, this process can be readily monitored by biochemical, antibody-based, and enzyme-based formats when the cell population examined is homogenous. When dealing with mixed and transfected cell populations however, such approaches may not be appropriate. Hence alternative methods are required. Here we describe flow-cytometry-based assays that can be utilized to examine signaling processes and degranulation in both pure mast cell populations and, following appropriate selection, in populations where the mast cells of interest may only represent a fraction of the total cell population.
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Abbreviations
- FcεRI:
-
High-affinity IgE receptor
- IP3 :
-
Inositol trisphosphate
- F actin:
-
Filamentous actin
- GFP:
-
Green fluorescent protein
- rhSCF:
-
Recombinant human stem cell factor
- dH2O:
-
Distilled or deionized H2O
- BSA:
-
Bovine serum albumin
- PFA:
-
Paraformaldehyde
- PBS:
-
Phosphate-buffered saline
- DNP:
-
Dinitrophenol
- HSA:
-
Human serum albumin
- APC:
-
Allophycocyanin
- DMSO:
-
Dimethylsulfoxide
- BMMC:
-
Bone marrow-derived mast cell
- FITC:
-
Fluorescein isothiocyanate
- PE:
-
Phycoerythrin
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Acknowledgement
Financial support for work in the authors’ laboratory was provided by the Division of Intramural Research of NIAID within the National Institutes of Health.
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Cruse, G., Gilfillan, A.M., Smrz, D. (2015). Flow Cytometry-Based Monitoring of Mast Cell Activation. In: Hughes, M., McNagny, K. (eds) Mast Cells. Methods in Molecular Biology, vol 1220. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1568-2_23
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DOI: https://doi.org/10.1007/978-1-4939-1568-2_23
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1567-5
Online ISBN: 978-1-4939-1568-2
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