This novel fluorescent probe enables the real-time detection of apoptosis without cell lysis or caspase activity interference. Comprising of a NucView® fluorogenic DNA dye attached to a DEVD substrate moiety, the NucView® caspase-3 substrates rapidly cross the cell membranes to enter the cytoplasm where cleavage by caspase-3 releases the high-affinity DNA dye. The released dye migrates to the cell nucleus, staining it which allows for visualisation of nuclear morphology in apoptotic cells by fluorescence microscopy. NucView® caspase-3 substrates are also compatible with flow cytometry and microscopy using a fluorescein filter set.
Benefits of using NucView® Caspase-3 substrates
• Detect caspase-3 activity real-time in live cells
• Enable visualisation of nuclear morphology real-time in live cells
• No caspase activity interference
• No cell lysis
• Simple protocol with no wash steps
• Fast protocol only requiring 15-30 minutes of incubation
• Non-toxic, compatible with multiday experiments
• Available with blue, green or red fluorescence
• Can be directly added into cell culture
• Compatible with adherent or suspension cells
• Compatible with flow cytometry and microscopy using a fluorescein filter set
• Validated in more than 100 cell types and 200 publications
"I'm using NucView® for a genome-wide siRNA screen looking at caspase3 activation. The NucView® is superior at detecting nuclear-specific caspase3 activity in the HCT116 cell line compared to standard caspase3 antibodies as it specifically localises in the nucleus with the Dapi staining"
Dr Nick Van Hateren, University of Sheffield
Staurosporine-induced MCF-7 Cells and NucView® 530
MCF-7 cells were left untreated or treated with staurosporine overnight to induce apoptosis, then stained for 30 minutes at 37⁰C with 40 nM MitoView™ Blue, 2 µM NucView® 530 Caspase-3 Substrate, and 100 ng/mL CF®488A Annexin V in cell culture medium. Healthy cells show mitochondrial staining with MitoView™ Blue (cyan), while apoptotic cells show NucView® 530 staining of nuclei (red) and CF®488A Annexin V staining of cell membranes (green).
Cell lines and primary cells tested with NucView® 488
To view the validated cell lines used with the NucView® 488 caspase-3 substrate with accompanying references, please click here
1. NucView® 405 Caspase-3 Substrate
NucView® 405 dye is excited by the 405 nm laser line for detection by confocal microscopy in the DAPI channel, or by flow cytometry in the Pacific Blue® channel. NucView® 405 is ideal for caspase-3 detection in multicolour applications.
2. NucView® 488 Caspase-3 Enzyme Substrate
The fluorescent staining produced in response to caspase-3/7 activity is compatible with subsequent fixation and permeabilisation for immunostaining. NucView® 488 Caspase-3 Substrate is offered as a 1 mM solution in DMSO or in PBS for use in cells that are sensitive to DMSO toxicity.
3. NucView® 530 Caspase-3 Substrate
NucView® 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence, for detection in the Cy®3 channel by fluorescence microscopy or the PE channel by flow cytometry. NucView® 530 can be used for multicolour imaging with blue, green, or far-red fluorescent probes. NucView® 530 Caspase-3 Substrate is offered as a 1 mM solution in DMSO or in PBS for use in cells that are sensitive to DMSO toxicity.
4. NucView® 488 and MitoViewTM 633 Apoptosis Assay Kit
The kit provides a convenient tool for profiling apoptotic cells based on caspase-3 activity and changes in the mitochondrial membrane potential using either fluorescence microscopy or flow cytometry.
5. NucView® 488 and RedDotTM 2 Apoptosis and Necrosis Kit
This kit provides a convenient tool for profiling apoptotic and necrotic cell populations by fluorescence microscopy or flow cytometry. The far-red dead-cell stain RedDot™ is included for staining necrotic and late apoptotic cells that have compromised plasma membrane integrity.
6. Dual Apoptosis Assay with NucView® 488 Caspase-3 Substrate and Annexin V
This kit provides two apoptosis markers, NucView® 488 Caspase-3 Substrate and Annexin V conjugated with a dye, for profiling apoptotic cells and studying the relation of two important apoptosis events, caspase-3 activation and phosphatidylserine (PS) translocation. Annexin V conjugation for the second marker is available with the following dyes: Sulforhodamine 101 (Texas Red®) Annexin V, CF®594 Annexin V, and CF®640R Annexin V.
Material available for download
NucView™ 488 Caspase-3 Assay Kit for Live Cells Protocol
NucView™488 and RedDot™2 Apoptosis & Necrosis Kit Protocol
NucView™ 488 and MitoView™ 633 Apoptosis Assay Kit Protocol
NucView™488 and CF™594 Annexin V Dual Apoptosis Assay Kit for Live Cells Protocol
NucView™ 488 Caspase-3 Substrate Protocol
Assays for Cell Viability & Cell Death
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