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Immunology and Infection

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JoVE Journal Immunology and Infection
Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry
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Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry

Article DOI: 10.3791/64526-v • 10:01 min • December 16th, 2022
December 16th, 2022 •
1, 1, 1
1Department of Immunology and Microbiology, University of Colorado-Anschutz School of Medicine

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Summary
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Calcium influx, a measure of T-cell signaling, is an effective way to analyze responses to T-cell receptor stimulation. This protocol for multiplexing Indo-1 with panels of antibodies directed at cell surface molecules takes advantage of the highly flexible capabilities of full spectrum flow cytometry.

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T-cell Receptor Calcium Influx Flow Cytometry Primary T-cells Subset Surface Phenotype Protocol Indo-1 AM DMSO CRPMI Cell Suspension Experimental Condition Single Stained Control Calcium-free Indo-1 EGTA Biological Negative Control Fluorophores Ionomycin Antibody Conjugated Fluorophores FC Receptor Blocking Antibody
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