NucView 530 Caspase-3 Substrate, 1 mM in DMSO



Product Description

NucView™ 530 Caspase-3 Substrate, 1 mM in DMSO, provides a convenient tool for detecting apoptosis in intact cells based on caspase-3/7 activity using either confocal microscopy or flow cytometry.

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. 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. The staining is also formaldehyde-fixable.

NucView™ 530 Caspase-3 Substrate stains apoptotic cell nuclei with orange fluorescence (Ex/Em 528/563 nm, Figure 1), for detection in the Cy®3 channel by fluorescence microscopy (Figure 2) or the PE channel by flow cytometry (Figure 3). 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.

Biotium also offers 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.


MitoView Blue, NucView 530, CF488A Annexin MCF7 composite
Figure 2. Detection of mitochondrial membrane potential, caspase-3 activity, and phosphatidylserine in MCF-7 cells. Cells were either untreated (A) or treated with staurosporine overnight to induce apoptosis (B), then stained with MitoView™ Blue, NucView™ 530 Caspase-3 Substrate, and CF™488A Annexin V for 30 minutes at 37oC in cell culture medium with no wash. Healthy cells show mitochondrial staining with MitoView Blue (cyan). Apoptotic cells lose MitoView™ Blue staining (cyan), and show NucView™ 520 staining of nuclei (red) and CF™488A Annexin V staining of cell membranes (green).


NucView 530, CF405M Annexin V staurosporine dose curve
Figure 3. Jurkat cells were treated with DMSO (negative control, blue) or 0.1 uM, 0.25 uM, or 1 uM staurosporine (red) to induce apoptosis in the presence of NucView™ 530 and CF™405M Annexin V for four hours at 37oC before analysis on a BD LSR II flow cytometer. Staining with NucView™ 530 and CF™405M Annexin V increase with increasing concentration of staurosporine.

Cy dye is a registered trademark of GE Healthcare.

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Please note that products will be shipped at ambient temperature. This will not affect performance if the products are kept at the indicated temperature for long term storage. Please refer to the package insert for correct storage conditions.

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