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NucView® 488 Caspase-3 Assay Kit for Live Cells

Green fluorescent caspase-3/7 substrate for detecting apoptosis in intact cells by confocal microscopy, flow cytometry, or real-time live cell imaging.

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, Homogeneous assay

Detection method/readout

Fluorescence microscopy, Live cell imaging, Flow cytometry

Substrate specificity

Caspases

Colors

Green

Fixation options

Fix after staining (formaldehyde), Permeabilize after staining

Excitation/Emission

500/530 nm (with DNA)

Storage Conditions

Store at 2 to 8 °C, Protect from light

Note: Due to operating restrictions related to our COVID-19 response, primary antibody conjugates of R-PE, APC, PerCP, HRP, or AP are temporarily unavailable. All other antibody options are still available. We apologize for the inconvenience. Please see our easy-to-use Mix-n-Stain™ Antibody Labeling Kits for labeling antibodies with R-PE, APC, tandem dyes, and enzymes.
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100 assays
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Product Description

NucView® 488 Caspase-3 substrate is a novel cell membrane-permeable fluorogenic caspase substrate designed for detecting caspase-3 activity within live cells in real time. See a video of NucView® 488 in action.

Features

  • Rapid, no-wash, endpoint or real-time assays
  • Non-toxic, allowing multi-day experiments to be performed
  • For flow cytometry, microscopy or live cell imaging systems
  • Dual detection of caspase activity and nuclear morphology
  • Formaldehyde fixable

Kit Components

  • NucView® 488 Caspase-3 substrate, 0.2 mM in DMSO
  • Caspase-3 inhibitor Ac-DEVD-CHO, 2 mM in DMSO

Spectral Properties

  • Ex/Em 500/530 nm (after cleavage, with DNA)

Unlike conventional caspase assays, NucView® 488 Caspase-3 substrate detects caspase-3 activity within individual intact cells without cell lysis or inhibition of caspase-3 activity. The substrate consists of a fluorogenic DNA dye and a DEVD substrate moiety specific for caspase-3. The substrate, which is both non-fluorescent and nonfunctional as a DNA dye, rapidly crosses cell membranes to enter the cytoplasm, where it is cleaved by caspase-3 to form a high-affinity DNA dye that stains the nucleus bright green. Thus, the NucView® 488 caspase-3 substrate is bi-functional, allowing detection of intracellular caspase-3 activity and visualization of changes in nuclear morphology during apoptosis. The fluorescent staining produced in response to caspase-3 activity is compatible with subsequent fixation and permeabilization for immunostaining.

The kit contains NucView® 488 Caspase-3 substrate and a caspase-3 inhibitor Ac-DEVD-CHO. The number of assays per kit is based on 5 uM substrate concentration in 200 uL assay volume; actual number of assays may vary based on concentration and staining volume used.

We also offer NucView® 488 Caspase-3 Substrate at 1 mM in DMSO (10402) or 1 mM in PBS (10403), for customers who wish to control the concentration of DMSO in their assay. Note that in cell types that are not sensitive to DMSO, the presence of DMSO during substrate incubation can enhance signal. See our full selection of NucView® substrates and kits.

NucView® enzyme substrate technology is covered by U.S. Patents.

References

 

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