Real Time Caspase-3 Activity Detection in Apoptotic Cells
Apoptotic cells within the same cell population can differ in the onset times for different apoptotic events. Therefore, it is important to be able to follow these events independently in real-time so that the interrelations among the events can be better understood. Traditionally, fluorescently-labeled caspase inhibitor assays (FLICA) have used irreversible inhibitors to label active caspases.
Cambridge Bioscience offers NucView™ 488 from Biotium. NucView™ 488 caspase-3 substrate is a novel fluorescent probe that does not interfere with caspase activity enabling the real-time detection of caspase-3 activity and visualization of nuclear morphology in apoptotic cells.
Substrate Design & Activity
The NucView™ 488 caspase-3 substrate is comprised of the NucView™ 488 DNA dye attached to the highly negatively charged caspase-3 substrate peptide sequence DEVD. Once linked to the substrate peptide, the DNA dye is unable to bind to DNA and remains non-fluorescent. This substrate can cross the plasma membrane to enter the cytoplasm, where it can be cleaved by caspase-3 to release the high-affinity DNA dye. This dye migrates to the cell nucleus to stain the nucleus with bright green fluorescence.
Benefits of Using NucView™ 488
• Does not interfere with caspase activity
• Detects caspase-3 activity real time in live cells
• Enables visualisation of nuclear morphology real time in live cells
• Enables fixable staining
• Can be directly added into cell culture
• Requires only 15-30 minutes of incubation with no wash
• Compatible with flow cytometry and microscopy using a fluorescein filter set
Cell Lines & Primary Cells Tested with NucView™ 488
The detection of caspase-3 activity using NucView™ 488 caspase-3 substrate has been reported for a wide variety of immortalized and primary cells in adherent, suspension and 3-D cultures as illustrated below:
To view the immortalized cell lines used with the Nucview™ 488 caspase-3 substrate with accompanying references, please Click Here
Formats Available
NucView™ 488 is available in the following kits and formats:
1. NucView™ 488 Caspase-3 Assay Kit for Live Cells
Contains NucView™ 488 Caspase-3 substrate in DMSO and caspase-3 inhibitor Ac-DEVD-CHO.
2. NucView™ 488 and MitoView™ 633 Apoptosis Kit
Includes Biotium's proprietary far red fluorescent MitoView 633 mitochondrial membrane potential dye (Ex/Em 622/648 nm) for simultaneous measurement of caspase-3 activity and mitochondrial membrane potential
3. Dual Apoptosis Assay with NucView™ 488 Caspase-3 Substrate& CF™594 Annexin V
Contains deep red fluorescent CF594 Annexin V (Ex/Em 593/614 nm) for dual detection of caspase-3 activity and phosphatidylserine translocation in intact cells (Figure 3). CF594 is brighter and more photostable than the spectrally similar dye Texas Red®.
4. NucView™ 488 Caspase-3 Enzyme Substrate
Substrate is offered as a 1 mM stock solution in DMSO or PBS. DMSO facilitates NucView™ 488 staining in some cell types. The substrate is offered in PBS for use in DMSO-sensitive cell types.
Apoptosis, Necrosis & Cell Viability Assays
To download the new Apoptosis, Necrosis & Cell Viability Assays brochure (PDF), please Click Here
Products
Note: product availability depends on country. Product detail pages show availability of each product.
