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

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

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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

Staining fixability

Fix/permeabilize after staining (formaldehyde/detergent)

Substrate specificity

Caspases

Colors

Orange

Size
Catalog #
price
Qty
Trial size (10 uL)
100 uL
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Product Description

NucView® 530 Caspase-3 Substrate, 1 mM in PBS, 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 Caspase-3 Substrate is available as a 1 mM solution in DMSO (catalog no. 10406) 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® 530 staining.

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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Support & Faq

NucView® Caspase 3 Enzyme Substrates

The substrates are very stable. Some users have reported performing time course assays with NucView® 488 Caspase-3 Substrate for 4-5 days.

Category: NucView® Caspase 3 Enzyme Substrates

← FAQs

NucView® Caspase-3 Substrates can be added to the cells at the start of the experiment or at the end. A major advantage of NucView® Caspase-3 Substrates compared to other apoptosis assays is that it can be used to monitor capase-3 activity in real time.

Category: NucView® Caspase 3 Enzyme Substrates

← FAQs

Yes. NucView® Caspase-3 Substrates are compatible with flow cytometers and other instruments with the appropriate excitation and emission settings.

Category: NucView® Caspase 3 Enzyme Substrates

← FAQs

← FAQs

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Shipping/Shelf-life

Shipping & Handling Disclaimer: Shipping and handling methods are calculated based on item temperature storage recommendation. Large, multi-item orders may be assessed additional fees. Shipping to Puerto Rico may be assessed additional fees. A Biotium customer service representative will contact you if additional fees are required. For expedited shipping, please request in the Order Notes section of the checkout page. Please note that products with recommended storage at 4°C or -20°C may ship at ambient temperature. This will not affect product performance. When you receive the product, place it under the recommended storage conditions.