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.
The CF® dye TUNEL Assay Apoptosis Detection Kit employs dUTP conjugated to the exceptionally bright and photostable fluorescent dyes, for single step fluorescent TUNEL labeling of fixed cells or tissues for analysis by fluorescence microscopy or flow cytometry.
The Apoptosis & Necrosis Quantitation Kit Plus provides a convenient assay for quantifying apoptotic (green) and necrotic (red) cells within the same cell population by flow cytometry or fluorescence microscopy
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.
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.