Microplate Assays for Caspase Activity

A distinctive feature of the early stages of apoptosis is the activation of caspase enzymes, which participate in the cleavage of protein substrates and in the subsequent disassembly of the cell. We offer a series of caspase assays that allow the simple detection of active caspases in living cells in real time or in cellular lysates or extracts.

Live-cell assays

CellEvent Caspase-3/7 Green Detection Reagent is optimized for apoptosis analysis with a simple no-wash protocol that helps preserve delicate apoptotic cells. Live cells in microplate wells can be monitored in real time for apoptotic activity by measuring fluorescence intensity.

Caspase-3/7 assays

Microplate assay kits for caspase-3/7 measurement provide a choice of substrates with different wavelength emissions. Kits are available using rhodamine 110 or AMC to report caspase activity. Substrates are also available as bulk reagents or predispensed into microplate wells

Dose–response curve with CellEvent Caspase-3/7 Green Detection Reagent. The percent positive for active caspase3/7 and EC50 were determined for staurosporine-treated U2OS cells using the reagents in the CellEvent Caspase-3/7 Green Detection Reagent.

Selection guide for microplate caspase assays

	CellEvent Caspase-3/7 Green Detection Reagent		EnzChek Caspase-3 Assay Kit #1, Z-DEVD-AMC Substrate	EnzChek Caspase-3 Assay Kit #2, Z-DEVD-R110 Substrate
Reporter	DEVD conjugated to a nucleic acid-binding dye		Z-DEVD-AMC substrate	Z-DEVD-R110 substrate
Ex/Em (nm)	502/530		342/441	496/520
Live cell	Yes		No	No
Lysate	No		Yes	Yes
Enzyme prep	No		Yes	Yes
Components	Enzyme substrate for live and fixed cells		Includes reference standard and inhibitor	Includes reference standard and inhibitor
For use with (Equipment)	Fluorescence microscope, High-content instrument, Microplate readers		Microplate readers	Microplate readers
Protocol outline	Add diluted CellEvent Caspase-3/7 Green Detection Reagent to cells Incubate for 30 minutes Measure fluorescence		Lyse cells, isolate supernatant. Prepare DEVD substrate and apply to wells. Add samples to wells. Incubate samples. Measure fluorescence. Determine caspase activity using standard curve.	Lyse cells, isolate supernatant Prepare DEVD substrate and apply to wells. Add samples to wells. Incubate samples. Measure fluorescence. Determine caspase activity using standard curve.
Format	25 µL	100 µL	500 assays using 100 µL reaction volume	500 assays using 100 µL reaction volume
Cat. No.	C10723	C10423	E13183	E13184

Expertly detect fluorescence with Thermo Scientific plate readers

High-sensitivity fluorescence detection for 96-1,536 samples can be quickly performed on Fluoroskan or Fluoroskan FL Microplate Fluorometer or Varioskan LUX Multimode Microplate Reader using Invitrogen reagents for optimal detection. Take advantage of automatic dynamic range selection to get optimal gain settings for each individual well and automation capabilities for even higher throughput.

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