Nucleofection of Caspase BiFC Components into Human Monocyte-Derived Macrophages
4:41
Treatment of Transfected MDM and Caspase BiFC Data Acquisition
6:37
Results: Differentiation of CD14-Monocytes and Transfection of Human Monocyte-Derived Macrophages
8:12
Conclusion
Transcript
This protocol is significant because it can be used to visualize and interrogate inflammatory caspase pathway at the molecular level in living cells in a particular disease state. The main advantage of this technique is that it can be used to iden
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This protocol describes the workflow to obtain monocytes-derived macrophages (MDM) from human blood samples, a simple method to efficiently introduce inflammatory caspase Bimolecular Fluorescence Complementation (BiFC) reporters into human MDM without compromising cell viability and behavior, and an imaging-based approach to measure inflammatory caspase activation in living cells.