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NucView 488 and RedDot 2 Apoptosis and Necrosis Kit

NucView™488 and RedDot™2 Apoptosis/Necrosis Assay Kit contains NucView™488 Caspase-3 Substrate for detection of caspase-3/7 activity. The far-red dead-cell stain RedDot™2 is included for staining necrotic and late apoptotic cells that have compromised plasma membrane integrity.

NucView 488
Catalog #

30072

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100 assays

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Product Description

NucView™488 and RedDot™2 Apoptosis/Necrosis Assay Kit contains NucView™488 Caspase-3 Substrate for detection of caspase-3/7 activity. The far-red dead-cell stain RedDot™2 is included for staining necrotic and late apoptotic cells that have compromised plasma membrane integrity. This kit provides a convenient tool for profiling apoptotic and necrotic cell populations by fluorescence microscopy or flow cytometry.

In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays, NucView™488 Caspase-3 Substrate can be used to detect caspase-3/7 activity within individual intact cells without inhibiting apoptosis progression. The substrate consists 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 bright green fluorescence. Thus, NucView™488 Caspase-3 Substrate allows detection caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis.

RedDot™2 is a cell membrane-impermeable, far-red dye with high selectivity for membrane compromised or dead cells. RedDot™2 has far-red emission at 695 nm, well-separated from the green fluorescence of NucView™488. The excitation maximum of RedDot™2 is 665 nm, but it can be efficiently excited by wavelengths from 488 to 647 nm, and therefore can be used with the 488 nm flow cytometry laser line.

Note: while NucView™488 staining is formaldehyde-fixable and compatible with subsequent immunostaining, fixation after staining with RedDot™2 is not recommended because it can result in increased background staining of healthy cells.

See our full selection of NucView™ substrates and kits.

NucView™ enzyme substrate technology is covered by U.S. patents.

 

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Support & Faq

NucView™ Caspase 3 Enzyme Substrates

The substrate is 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

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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

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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

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Apoptosis, Necrosis, and Cell Viability Kits

MTT and XTT are colorimetric based assays, while resazurin can be measured using colorimetric or fluorescence detection. MTT is not a soluble product, so the cells must be lysed to solubilize the formazan salt before absorbance can be measured. XTT and resazurin do not require cell lysis, allowing kinetic monitoring of the same samples at different timepoints.

Category: Apoptosis, Necrosis, and Cell Viability Kits

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Check to make sure your cell densities are in the linear range of the cell viability assay. Too few cells may fall below the limit of detection, and too many cells may saturate the OD. Perform an initial experiment to check the best linear range for your cell type and experimental set-up. You may need to vary the cell density assay incubation time. The kit protocols provide general guidelines and may need to be optimized for your experimental system.

Category: Apoptosis, Necrosis, and Cell Viability Kits

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Resazurin is reduced by cells to the fluorescent product resorufin. Resorufin can be reduced further to a non-fluorescent compound. Therefore the densest wells may have lowest fluorescence due to over-reduction of the substrate. Please see the product information sheet for more details. The kit protocol provides general guidelines and may need to be optimized empirically for your experimental system. You may need to vary cell density or assay incubation time to ensure that your samples fall in the linear range of the kit.

Category: Apoptosis, Necrosis, and Cell Viability Kits

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Shipping/Shelf-life

Shipment Method: Shipping and handling methods will be assessed and calculated at time of shipment based upon item(s) storage temperature conditions. Expedited shipment may be requested at time of checkout. 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.