This in vitro assay employs the far-red fluorescent inhibitor probe 660-DEVD-FMK to label active caspase 3 and caspase 7 enzymes in living cells. Analyze the fluorescent signal using fluorescence microscopy or flow cytometry.
Samples:
Cell culture
Application Notes:
Application Notes: 1. Prepare Assay buffer, dilute Assay Buffer Concentrate 1:5 in diH2O., 2. Prepare samples, dilute > 1:10 in Assay Buffer., 3. Prepare standard curve and zero (blank) standard., 4. Add 50 µL blanks, standards, and samples to plate., 5. Add 25 µL Fluorescent H2O2 Detection Substrate to plate., 6. Prepare HRP, dilute Horseradish Peroxidase Concentrate 1:100 in Assay Buffer., 7. Add 25 µL HRP Preparation to plate., 8. Incubate 15 minutes at room temperature., 9. Read fluorescence with excitation at 570 nm and emission at 590 nm
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