Name: NucView® 530 Caspase-3 Substrate, 1 mM in DMSO
SKU: 10406
Availability: InStock

Biotium products are distributed only in Singapore and Thailand.

NucView® 530 Caspase-3 Substrate detects apoptosis in intact cells based on caspase-3/7 activity using confocal microscopy, flow cytometry, or live cell imaging.

NucView® 530 dye has excitation/emission maxima of 528/563 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 detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. Staining is compatible with subsequent fixation and permeabilization for immunostaining.

NucView® 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence (Ex/Em 528/563 nm), for detection in the Cy®3 channel by fluorescence microscopy or the PE channel by flow cytometry. NucView® 530 can be used for multi-color imaging with blue, green, or far-red fluorescent probes. Note that when excited by the 488 nm laser line, NucView® 530 also fluoresces in the FITC channel, and therefore cannot be analyzed together with green probes by flow cytometry.

NucView® 530 is also available as 1 mM in PBS (catalog number 10408) for use with DMSO-sensitive cell types.

We also offer blue fluorogenic NucView® 405 Caspase-3 Substrate, and green fluorogenic NucView® 488 Caspase-3 Substrate and kits.

Excitation and emission spectra of NucView 530 dye with dsDNA. — Figure 1. Excitation and emission spectra of NucView® 530 dye with dsDNA.

NucView® enzyme substrate technology is covered by U.S. patents. Cy dye is a registered trademark of GE Healthcare.

Reference Publications

Tang, Ho Man, and Peter Chi Keung Cheung

Gallic Acid Triggers Iron-Dependent Cell Death with Apoptotic, Ferroptotic, and Necroptotic Features

Toxins vol. 11,9 492.

DOI: 10.3390/toxins11090492

Article Snippet: “For detecting the caspase-3 activity using the NucView 530 Caspase-3 substrate (#10406) from Biotium (Fremont, CA, USA), the substrate was diluted with cell culture medium 1:2000, and then applied to the cells and incubated at 37 °C under an atmosphere of 5% CO₂ for 20 min before subjected to microscopy.”

Yusuke Mori, Naoka Shiratsuchi, et al.

Extracellular ATP facilitates cell extrusion from epithelial layers mediated by cell competition or apoptosis

Current Biology 32, 2144–2159

DOI: https://doi.org/10.1016/j.cub.2022.03.057

Article Snippet: “Nucview 530 Caspase-3 substrate (2 mM) (Biotium) was then added together with tetracycline and apyrase. After 2 h of the treatment, the activity of caspase-3 was analyzed by time-lapse imaging for 24 h.”

Merete Bridget Long

The role of neutrophil-derived microvesicles in lung inflammation

Department of Infection, Immunity and Cardiovascular Disease School of Medicine The University of Sheffield

DOI: Thesis_Merete Long

Article Snippet: Highlight from the Materials and suppliers table “NucView® 530 Caspase-3 Substrate Biotium 10406″

Minnah Thomas 1, Benoit Ladoux 2, Yusuke Toyama

Desmosomal Junctions Govern Tissue Integrity and Actomyosin Contractility in Apoptotic Cell Extrusion

Current Biology 30, 682–690

DOI: https://doi.org/10.1016/j.cub.2020.01.002

Article Snippet: Highlight from the Chemicals, Peptides, and Recombinant Proteins table “NucView 488 Caspase-3 assay kit Biotium Cat#30029, NucView 530 Caspase-3 assay Kit Biotium Cat#10406″

Luke, Cliff J et al.

Lysoptosis is an evolutionarily conserved cell death pathway moderated by intracellular serpins

Communications biology vol. 5,1 47.

DOI: 10.1038/s42003-021-02953-x

Article Snippet: “Acridine orange (AO) (40039) and NucView^® 530 (10406) were purchased from Biotium. FITC-annexin V was purchased from Southern Biotech or FITC-annexin V apoptosis detection kit (556547) was purchased from BD Pharmingen.”