For flow cytometry, microscopy, or live cell imaging.
Formaldehyde-fixable nuclear staining.
Green fluorescent NucView 488
Orange fluorescent NucView 530
NucView™ 488 Caspase-3 Enzyme Substrate in action. HT-1080 cells were treated with camptothecin and NucView™ 488 Caspase-3 Substrate and imaged every 30 minutes for 24 hours on the IncuCyte® Zoom. Apoptotic cell nuclei fluoresce green as NucView™ 488 dye is released by caspase-3 cleavage. Video courtesy of Essen Biosciences.
Principle of NucView™ caspase substrates
NucView™ Caspase-3 Substrates are novel fluorogenic substrates developed by Biotium to detect apoptosis in intact cells in real-time. Biotium has invented a new concept in the design of fluorogenic enzyme substrates. An enzyme substrate is attached to a non-functional fluorescent dye. Upon enzymatic cleavage of the substrate, the dye is released and becomes capable of binding to its target to emit fluorescence. In the case of NucView™ Caspase-3 Substrates, the caspase-3 substrate peptide DEVD is attached to a DNA-binding dye. Before cleavage, the dye is unable to bind to DNA and is thus non-fluorescent. The substrate enters the cell cytoplasm where it is cleaved by caspase-3 to release the fluorogenic DNA dye, which stains the nucleus.
Advantages of NucView™ caspase substrates
- Unlike FLICA assays that use irreversible inhibitors to label active caspases, NucView™ caspase-3 substrates do not interfere with caspase activity, allowing monitoring of caspase activity in live cells.
- Non-toxic, allowing for multi-day experiments.
- Bi-functional substrate allows one to both detect caspase-3 activity within an apoptotic cell but also to visualize apopototic nuclear morphology.
- Green fluorogenic NucView™ 488 Caspase-3 Substrate can be detected in the FITC channel by fluorescence microscopy, flow cytometry, or fluorescence microplate reader.
- Detection of apoptosis using NucView™ 488 Caspase-3 Substrate has been reported in more than 90 different primary and immortalized cells types, including 3-D cultures, in more than 150 publications.
NucView™ caspase-3 substrates
- The original green fluorogenic NucView™ 488 can be detected in the FITC channel by fluorescence microscopy, flow cytometry, or fluorescence microplate reader.
- The blue NucView™ 405 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, for multi-color applications in which the green fluorescence channel is reserved for other detection reagents.
- The orange NucView™ 530 has excitation/emission mexima of 528/563 nm, and can be detected by microscopy or flow cytometry in the Cy®3 or R-PE channels.
- NucView™ caspase-3 substrates are available as a stand-alone substrates, or in kits with other fluorescent apoptosis and necrosis probes for studying the temporal and spatial relations among the different events by fluorescence microscopy or flow cytometry.