NucView® 405 Caspase-3 Substrate, 1 mM in PBS

NucView® 405 Caspase-3 Substrate, 1 mM in PBS

NucView® 405 Caspase-3 Substrate, 1 mM in PBS

Add to cart

Blue fluorescent caspase-3/7 substrate for detecting apoptosis in intact cells by confocal microscopy, flow cytometry, or live cell imaging.

Product Description

NucView® 405 Caspase-3 Substrate is a convenient tool for detecting apoptosis in intact cells based on caspase-3/7 activity using confocal microscopy, flow cytometry, or live cell imaging.

  • NucView® 405 dye has excitation/emission at 429/469 nm (with DNA)
  • Rapid, no-wash, endpoint or real-time assays
  • Non-toxic, allowing multi-day experiments to be performed
  • For flow cytometry, microscopy or live cell imaging systems
  • Dual detection of caspase activity and nuclear morphology
  • Formaldehyde fixable

NucView® substrates consist of a fluorogenic DNA dye coupled to the caspase-3/7 DEVD recognition sequence. The substrate, which is initially non-fluorescent, penetrates the plasma membrane and enters the cytoplasm. In apoptotic cells, caspase-3/7 cleaves the substrate, releasing the high-affinity DNA dye, which migrates to the cell nucleus and stains DNA with fluorescence. Thus, NucView caspase-3 substrates are bifunctional, allowing detection of caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis. In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, NucView® caspase-3 substrates can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. Staining is compatible with subsequent fixation and permeabilization for immunostaining.

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 multi-color applications for researchers who wish to reserve the green fluorescence channel for other detection reagents.

Note: NucView® 405 signal may be difficult to observe using epifluorescence microscopy. Confocal microscopy using 405 nm laser excitation is recommended for imaging.

NucView® 405 Caspase-3 Substrate is offered as a 1 mM solution in DMSO (catalog no. 10405) or PBS (phosphate-buffered saline). The substrate in PBS is formulated for use in cells that are sensitive to DMSO toxicity. In non-DMSO sensitive cell types, adding DMSO during the substrate incubation may enhance NucView® 405 staining.

Note: NucView® 405 signal may be difficult to observe using epifluorescence microscopy. Confocal microscopy using 405 nm laser excitation is recommended for imaging. Biotium also offers green fluorescent NucView® 488 caspase-3 substrate and kits as well as orange fluorescent NucView® 530.

Pacific Blue is a registered trademark of Thermo Fisher Scientific.

Product Attributes

Apoptosis/viability marker Caspase
For live or fixed cells For live/intact cells
Assay type/options Long term staining (24-72h), Endpoint assay, No-wash staining, Real-time imaging, Homogeneous assay
Detection method/readout Fluorescence microscopy, Live cell imaging, Flow cytometry
Substrate specificity Caspases
Colors Blue
Fixation options Fix after staining (formaldehyde), Permeabilize after staining
Excitation/Emission 429/469 nm (with DNA)
Storage Conditions Store at -10 to -35 °C

Reference Publications

Ichimaru, Yoshimi et al.

Sasa veitchii extract induces anticancer effects via inhibition of cyclin D1 expression in MCF-7 cells

Nagoya journal of medical science vol. 82,3 (2020): 509-518.

DOI: 10.18999/nagjms.82.3.509

Article Snippet:To measure apoptotic cells, we used a Dual Apoptosis Assay Kit with NucView 405 Caspase-3 Substrate (Biotium Inc) and an Annexin V-FITC Detection kit (Nacalai Tesque) according to the manufacturer’s instructions.”

 

Kim, H., Watanabe, S., Kitamatsu, M. et al.

Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate

Sci Rep 10, 19087 (2020).

DOI: https://doi.org/10.1038/s41598-020-76100-7

Article Snippet:At 5 h after photoirradiation, the NucView405 Caspase-3 Substrate (Biotium, Fremont, CA, USA) was added to the medium at a final concentration of 100 μM to stain apoptotic cells.”

Documents