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NucView Caspase-3 Substrates

The Only Caspase Substrate for Real-Time Apoptosis Detection

Real-time apoptosis detection
Substrate doesn't interfere with apoptosis progression.
For flow cytometry, microscopy, or live cell imaging.
Fast, simple, and fixable
Homogenous, no wash 15-30 minute assay.
Formaldehyde-fixable nuclear staining.
Three color options
Blue fluorescent NucView 405
Green fluorescent NucView 488
Orange fluorescent NucView 530

Principle of NucView substrate technology. The substrate is non-fluorescent and does not bind DNA. After enzyme cleavage, the high-affinity DNA dye is released to bind DNA and become fluorescent.


NucView™ 488 Caspase-3 Enzyme Substrate in action. HT-1080 cells were treated with camptothecin and NucView™ 488 Caspase-3 Substrate and imaged every 30 minutes for 24 hours on the IncuCyte® Zoom. Apoptotic cell nuclei fluoresce green as NucView™ 488 dye is released by caspase-3 cleavage. Video courtesy of Essen Biosciences.

Principle of NucView™ caspase substrates

NucView™ Caspase-3 Substrates are novel fluorogenic substrates developed by Biotium to detect apoptosis in intact cells in real-time. Biotium has invented a new concept in the design of fluorogenic enzyme substrates. An enzyme substrate is attached to a non-functional fluorescent dye. Upon enzymatic cleavage of the substrate, the dye is released and becomes capable of binding to its target to emit fluorescence. In the case of NucView™ Caspase-3 Substrates, the caspase-3 substrate peptide DEVD is attached to a DNA-binding dye. Before cleavage, the dye is unable to bind to DNA and is thus non-fluorescent. The substrate enters the cell cytoplasm where it is cleaved by caspase-3 to release the fluorogenic DNA dye, which stains the nucleus.

Advantages of NucView™ caspase substrates

  • Unlike FLICA assays that use irreversible inhibitors to label active caspases, NucView™ caspase-3 substrates do not interfere with caspase activity, allowing monitoring of caspase activity in live cells.
  • Non-toxic, allowing for multi-day experiments.
  • Bi-functional substrate allows one to both detect caspase-3 activity within an apoptotic cell but also to visualize apopototic nuclear morphology.
  • Green fluorogenic NucView™ 488 Caspase-3 Substrate can be detected in the FITC channel by fluorescence microscopy, flow cytometry, or fluorescence microplate reader.
  • Detection of apoptosis using NucView™ 488 Caspase-3 Substrate has been reported in more than 90 different primary and immortalized cells types, including 3-D cultures, in more than 150 publications.

NucView™ caspase-3 substrates

  • The original green fluorogenic NucView™ 488 can be detected in the FITC channel by fluorescence microscopy, flow cytometry, or fluorescence microplate reader.
  • The blue NucView™ 405 is excited by the 405 nm laser line for detection by confocal microscopy in the DAPI channel, or by flow cytometry in the Pacific Blue® channel,  for multi-color applications in which the green fluorescence channel is reserved for other detection reagents.
  • The orange NucView™ 530 has excitation/emission mexima of 528/563 nm, and can be detected by microscopy or flow cytometry in the Cy®3 or R-PE channels.
  • NucView™ caspase-3 substrates are available as a stand-alone substrates, or in kits with other fluorescent apoptosis and necrosis probes for studying the temporal and spatial relations among the different events by fluorescence microscopy or flow cytometry.

Jurkat cells were treated with staurosporine to induce apoptosis 1-5 hours, stained with NucView and analyzed in FL1 of a BD FACSCalibur flow cytometer.

Time course of caspase-3 detection with NucView™ 405 Caspase-3 Substrate in Jurkat cells treated with staurosporine to induce apoptosis, analyzed in the Pacific Blue® channel of a BD LSRII flow cytometer (405 nm excitation, 450/50 nm emission filter).


MCF-7 cells were left untreated or treated with staurosporine overnight to induce apoptosis, then stained for 30 minutes at 37C with 40 nM MitoView Blue, 2 uM NucView 520 Caspase-3 Substrate, and 100 ng/mL CF488A Annexin V in cell culture medium. Healthy cells show mitochondrial staining with MitoView Blue (cyan), while apoptotic cells show NucView 520 staining of nuclei (red) and CF488A Annexin V staining of cell membranes (green).


Apoptotic Jurkat cell stained with NucView 488 (green) and CF647 Annexin V (magenta).

 

 

Dual Apoptosis Assay with NucView 488 Caspase-3 Substrate & Annexin V

From: $341 Sizes: 50 assaysCatalog #: 30030, 30067, - 30073View allHide

NucView 405 Caspase-3 Substrate, 1 mM in DMSO

From: $63 Sizes: Trial size (10 uL), 100 uLCatalog #: 10405, - 10405-TView allHide

NucView 488 and MitoView 633 Apoptosis Assay Kit

From: $341 Sizes: 100 assaysCatalog #: 30062

NucView 488 and RedDot 2 Apoptosis and Necrosis Kit

From: $299 Sizes: 100 assaysCatalog #: 30072

NucView 488 Caspase-3 Assay Kit for Live Cells

From: $92 Sizes: Trial size (25 assays), 100 assaysCatalog #: 30029, - 30029-TView allHide

NucView 488 Caspase-3 Enzyme Substrate, 1 mM in DMSO

From: $63 Sizes: 10 uL, 100 uLCatalog #: 10402, - 10402-TView allHide

NucView 488 Caspase-3 Enzyme Substrate, 1 mM in PBS

From: $63 Sizes: 10 uL, 100 uLCatalog #: 10403, - 10403-TView allHide

NucView 530 Caspase-3 Substrate, 1 mM in DMSO

From: $63 Sizes: Trial size (10 uL), 100 uLCatalog #: 10406, - 10406-TView allHide

 

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

← 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

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← FAQs

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