Indo-1 Ratiometric Dye and Fluorescent Antibody Labeling
3:35
Acquisition of Calcium Influx Using Flow Cytometric Analysis
7:45
Results: Visualization of Calcium Responses in Gated Populations of Murine Thymocytes
9:17
Conclusion
Transcription
The protocol allows investigators to simultaneously measure calcium response in primary T-cells along with assessing their subset and surface phenotype. The main advantage of this technique is that it avoids the need to isolate specific subsets of
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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.