NucView® 488 and RedDot™ 2 Apoptosis & Necrosis Kit (100 Assays)

Dual-fluorescence detection of apoptosis and necrosis using caspase-3/7 activity and membrane integrity markers.

Product Description

The NucView® 488 and RedDot™ 2 Apoptosis & Necrosis Kit is a dual-fluorescence assay designed to distinguish apoptotic and necrotic cell populations. It combines NucView® 488 Caspase-3 Substrate for detecting caspase-3/7 activity with RedDot™ 2, a far-red fluorescent dead-cell stain that identifies cells with compromised plasma membrane integrity.

This combination provides a convenient approach to investigating cell death pathways using fluorescence microscopy or flow cytometry. By monitoring caspase activation and membrane integrity simultaneously, researchers can characterize different stages of cell death within the same sample.

NucView® 488 uses a fluorogenic DNA dye linked to the DEVD recognition sequence of caspase-3/7. Initially non-fluorescent, the substrate enters intact cells and is cleaved by activated caspase-3/7 during apoptosis. The released fluorescent DNA-binding dye migrates into the nucleus, producing bright green fluorescence and allowing researchers to observe nuclear changes associated with apoptosis.

RedDot™ 2 is a membrane-impermeable far-red fluorescent dye that selectively stains dead or membrane-compromised cells. Its emission maximum is approximately 695 nm, providing spectral separation from the green fluorescence of NucView® 488. This makes the two dyes suitable for combined fluorescence-based analysis.

Assay Principle: How Does It Work?

The kit uses two complementary fluorescent indicators to assess cellular viability and death:

Combined analysis: Green and far-red fluorescence provide complementary information about caspase activation and loss of membrane integrity.

Key Features and Benefits

• Dual-fluorescence detection: Simultaneous assessment of apoptotic and membrane-compromised cells.
• Caspase-3/7 monitoring: Detects enzymatic activity associated with apoptosis.
• Green fluorescence: NucView® 488 provides clear nuclear staining in apoptotic cells.
• Far-red fluorescence: RedDot™ 2 identifies dead and membrane-compromised cells.
• Non-inhibitory detection approach: NucView® 488 detects caspase activity without inhibiting apoptosis progression.
• Multiple detection platforms: Suitable for fluorescence microscopy and flow cytometry.
• Cell morphology observation: Enables visualization of nuclear morphological changes during apoptosis.

Applications

The NucView® 488 and RedDot™ 2 Apoptosis & Necrosis Kit can support a variety of cell biology and biomedical research applications, including:

• Apoptosis and necrosis research.
• Cell death pathway analysis.
• Drug-induced cytotoxicity studies.
• Cell viability and cell death profiling.
• Fluorescence microscopy experiments.
• Flow cytometry analysis of cell populations.
• Evaluation of cellular responses to experimental treatments.
• Cancer biology and cell signaling research.

Technical Specifications

Important Technical Notes

• NucView® 488 staining is formaldehyde-fixable and compatible with subsequent immunostaining.
• Fixation after RedDot™ 2 staining is not recommended because it may increase background staining in healthy cells.
• RedDot™ 2 emits in the far-red region, providing spectral separation from NucView® 488 green fluorescence.
• For optimal results, follow the manufacturer's recommended staining and handling instructions.

Storage and Stability

The reported product stability and shelf life are both 6 months. Refer to the manufacturer's product documentation for the complete storage temperature, handling requirements, and reagent preparation instructions.

Explore Apoptosis and Necrosis Research Solutions

Support your cell death analysis with fluorescence-based research tools for apoptosis and necrosis detection.