Cell Meter™ Live Cell Caspase 3/7 Imaging Kit *Green Fluorescence*
|Shipping||Standard overnight for United States, inquire for international|
|Extinction coefficient (cm -1 M -1)||80000|
|H-phrase||H303, H313, H333|
|Intended use||Research Use Only (RUO)|
|R-phrase||R20, R21, R22|
Extinction coefficient (cm -1 M -1)
|Excitation||FITC filter set|
|Emission||FITC filter set|
|Recommended plate||Black wall/clear bottom|
AT A GLANCE
- Prepare cells with test compounds at a density of 5 × 104 to 2 × 105 cells/100 µL/well/96-well plate
- Add equal volume of Caspase 3/7 Substrate working solution
- Incubate in a 5% CO2 incubator at 37 °C for 60 minutes
- Image with a fluorescence microscope using a FITC filter set
Thaw all the kit components at room temperature before use.
For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 50 µL of DMSO (Component C) into the vial of ApoSight™ Green Caspase 3/7 Substrate (Component A) to make 200X ApoSight™ Green Caspase 3/7 Substrate stock solution.
Note Aliquot in smaller vials to avoid repeated freeze thaw cycles. Protect from light.
PREPARATION OF WORKING SOLUTION
Prepare ApoSight™ Green Caspase 3/7 substrate working solution by mixing 5 µL of 200X ApoSight™ Green Caspase 3/7 Substrate stock solution with 1 mL of assay buffer (Component B). Mix well.
Note 100 µL of ApoSight™ Green Caspase 3/7 substrate working solution is enough for 10 tests in a 96-well plate format
Note Prepare enough ApoSight™ Green Caspase 3/7 substrate working solution right before the experiment, and use promptly.
SAMPLE EXPERIMENTAL PROTOCOL
Treat Jurkat cells with 2 μg/mL camptothecin for 3 hours
Treat Jurkat cells with 1 μM staurosporine for 3-4 hours
Treat HL-60 cells with 4 μg/mL camptothecin for 4 hours
Treat HL-60 cells with 1 μM staurosporine for 4 hours.
Note Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.
- Prepare cells with test compounds at a density of 5 × 104 to 2 × 105 cells/100 µL/well/96-well plate.
- Add equal volume of Caspase 3/7 Substrate working solution to the cells (100 µL/well/96 well-plate).
Incubate in a 5% CO2 incubator at 37 °C for 60 minutes.
Image with a fluorescence microscope using a FITC filter set.
|Extinction coefficient (cm -1 M -1)||80000|
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