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NucView® 530 Caspase-3 Substrate, 1 mM in DMSO

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.

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, Homogenous assay

Detection method/readout

Fluorescence microscopy, Live cell imaging, Flow cytometry

Substrate specificity

Caspases

Colors

Orange

Fixation options

Fix after staining (formaldehyde), Permeabilize after staining

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Catalog #
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Trial size (10 uL)
100 uL
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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.

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