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The Only Caspase Substrates for Real-Time Apoptosis Detection

NucView® caspase-3 substrates are Biotium’s patented fluorogenic substrates designed for real-time apoptosis detection in live cells.

Real-time apoptosis detection
Substrate doesn't interfere with apoptosis progression.
For flow cytometry, microscopy, or live cell imaging.
Fast, simple, and fixable
Homogeneous, 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® caspase-3 substrates are based on novel fluorogenic DNA dyes that have been coupled to the caspase-3/7 recognition sequence (DEVD). When introduced to cells, the substrate is initially non-fluorescent and is able to penetrate the plasma membrane and enter the cytoplasm. During apoptosis, caspase-3/7 cleaves the substrate and releases the high-affinity DNA dye which leads to nuclear fluorescent staining. Consequently, NucView caspase-3 substrates are bifunctional, allowing detection of caspase-3/7 activity and visualization of morphological changes in the nucleus during apoptosis.

In contrast to other fluorogenic caspase substrates or fluorescent caspase inhibitor based (FLICA) assays that which inhibit caspase activity, NucView® caspase-3 substrates can be used to detect caspase-3/7 activity in live cells without inhibiting apoptosis progression. Staining is also compatible with subsequent fixation and permeabilization for downstream immunostaining.


  • Dual detection of caspase activity and nuclear morphology
  • Rapid, no-wash, endpoint or real-time assays
  • Non-toxic, allowing for multi-day experiments
  • For flow cytometry, microscopy, or live cell imaging systems
  • Available as stand-alone substrates or in kits paired with other apoptosis or necrosis probes
Click to view GIF animation. HeLa cells were stained with NucView® 488 Caspase-3 Substrate (2 uM), MitoView™ 633 mitochondrial potential dye (50 nM), and Hoechst 33342 blue nuclear stain (1 ug/mL), then treated with 1 uM staurosporine to induce apoptosis.

Available in Blue, Green, or Red Fluorescence

NucView® 405 Caspase-3 Substrate, 1 mM in PBS
Apoptotic HeLa cells stained with NucView® 405 Caspase-3 Substrate (blue) and CF®488A Annexin V (green).
Apoptotic HeLa cells stained with NucView® 488 Caspase-3 Substrate (green) and CF®594 Annexin V (red).
NucView 530 CF488A Annexin
Apoptotic MCF-7 cells stained with NucView® 530 Caspase-3 Substrate (red) and CF®488A Annexin V (green).

Analyze Apoptosis by Flow Cytometry in Live Cells

Jurkat cells were treated with DMSO (negative control, blue) or 0.1 uM, 0.25 uM, or 1 uM staurosporine (red) to induce apoptosis in the presence of NucView® 530 and CF®405M Annexin V for four hours at 37°C before analysis on a BD LSR II flow cytometer. Staining with NucView® 530 and CF®405M Annexin V increase with increasing concentration of staurosporine.
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).

Dual Detection Kits

Explore assay kits that combine the advantage of NucView® real-time apoptosis analysis with other probes for monitoring necrosis, membrane potential, or other apoptosis markers.


Flow cytometry analysis of Jurkat cells treated with staurosporine (green) to induce apoptosis, or DMSO controls (pink), using the NucView® 488 and MitoView™ 633 Apoptosis Kit. Fluorescence was analyzed on a BD FACSCalibur flow cytometer. As apoptosis progresses over time in staurosporine-treated cells, NucView® 488 signal (FL1, x-axis) increases and mitochondrial membrane potential measured by MitoView™ 633 staining (FL4, y-axis) decreases.

NucView® Caspase-3 Substrates and Kits

Catalog No.


NucView® 405 Caspase-3 Substrate, 1 mM in DMSO10405Blue fluorescence for flow cytometry in the Pacific Blue® channel or microscopy with the 405 nm laser
NucView® 405 Caspase-3 Substrate, 1 mM in PBS10407NucView® 405 substrate in PBS, for DMSO-sensitive cell types
NucView® 488 Caspase-3 Substrate, 1 mM in DMSO10402Green fluorescent substrate validated in more than 100 cell types and 200 publications
NucView® 488 Caspase-3 Substrate, 1 mM in PBS10403NucView® 488 substrate in PBS, for DMSO-sensitive cell types
NucView® 530 Caspase-3 Substrate, 1 mM in DMSO10406Orange fluorescence for microscopy in the Cy®3 channel or flow cytometry in the R-PE channel
NucView® 530 Caspase-3 Substrate, 1 mM in PBS10408NucView® 530 substrate in PBS, for DMSO-sensitive cell types
NucView® 488 and MitoView™ 633 Apotosis Detection Kit30062NucView® 488 and far-red fluorescent MitoView™ 633 for the Cy®5 channel
NucView® 488 and RedDot™ 2 Apoptosis & Necrosis Kit30072NucView® 488 and far-red dead cell DNA dye RedDot™ 2 for the Cy®5 channel
Dual Apoptosis Assay with NucView® 488 and CF®594 Annexin V30067NucView® 488 and red fluorescent Annexin V apoptosis probe
Dual Apoptosis Assay with NucView® 488 and CF®640R Annexin V30073NucView® 488 and far-red fluorescent Annexin V apoptosis probe


NucView® 488 is Validated in More Than 100 Cell Types and 200 Publications

Highlighted Citation: NucView® 488 was used in a T cell-mediated tumor cell killing assay by Lim et al. in a study investigating the role of inflammation on PD-L1 stabilization. Results show T-cell mediated killing of PD-L1 expressing BT549 ductal carcinoma cells was no longer suppressed when treated with neutralizing TNF-α antibody. In addition, PD-L1 depleted cells did not respond to either cytokine enriched macrophage-conditioned medium (MP) or TNF-α antibody, suggesting that PD-L1 expression is required for TNF- α-mediated suppression of T cell activity.

See the full list of references here.

T cell-meditated tumor cell killing assay in PD-L1 knockout (PD-L1 KO) BT549 cells. Representative phase, red fluorescent (nuclear restricted RFP), and green fluorescent (NucView® 488 caspase-3/7 substrate) merged images of activated T cell co-cultures in the presence of caspase-3/7 substrate at 96 hr are shown. T cells were activated with CD3 antibody (100 ng/mL) and IL-2 (10 ng/mL). Green fluorescent cells were counted as dead cells. The quantitative ratio of dead cells is shown in the bar graph on the right. Scale bar, 100 um. Credit: Lim et al.

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

From: $369 Sizes: 50 assaysCatalog #: 30030, 30067, - 30073View allHide

NucView® Caspase-3 Enzyme Substrates

From: $68 Sizes: 10 uL (1 mM in DMSO), 100 uL (1 mM in DMSO), 10 uL (1 mM in PBS), 100 uL (1 mM in PBS)Catalog #: 10405-T, 10405, 10407-T, 10407, 10402-T, 10402, 10403-T, 10403, 10406-T, 10406, 10408-T, - 10408View allHide

NucView® 488 & MitoView™ 633 Apoptosis Assay Kit

From: $369 Sizes: 100 assaysCatalog #: 30062

NucView® 488 & RedDot™ 2 Apoptosis & Necrosis Kit

From: $323 Sizes: 100 assaysCatalog #: 30072

NucView® 488 Caspase-3 Assay Kit for Live Cells

From: $99 Sizes: Trial size (25 assays), 100 assaysCatalog #: 30029, - 30029-TView allHide



NucView® Caspase-3 Enzyme Substrates

Like other caspase-3 substrates, NucView® 488 Caspase-3 Substrate is based on a DEVD sequence that also can be cleaved by caspase-7.

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The tables below list primary cells types and immortalized cell lines reported to work with NucView® 488 in the published scientific literature. Click here for a PDF version of the tables along with the references.
Primary cell typeSpecies
Alveolar epithelial cellsMouse
Cortical neuronsRat
Dendritic cellsMouse
Embryonic fibroblast (MEF)Mouse
Embryo tailbudChicken
Gingival fibroblastsHuman
HemocytesSilkworm(Bombyx mori)
Hippocampal neuronsRat
Idiopathic pulmonary fibrosis fibroblastsHuman
Immature B cellsMouse
Kidney epithelial cellsMouse
Lung microvascular endothelial cellsHuman
Mammary epithelial cells (3-D cultures)Mouse
SVZ neural progenitor cellsRat
OocytesBovine, mouse
Pancreatic acinar cellsMouse
Pancreatic beta cellsRat
Pancreatic islet cellsMouse
Peritoneal macrophagesMouse
Pollen tubesField poppy(Papaver rhoeas)
Retinal pigmented epithelial cellsHuman, mouse
Skin fibroblastsSand cat(Felis margarita)
Stem cellsHuman
Umbilical vein endothelial cellsHuman
Vascular endothelial cellsRat
Immortalized cell lineSpeciesCell type
293-HHumanEmbryonic kidney
293-THumanEmbryonic kidney
4T1MouseMammary tumor
67NRMouseMammary carcinoma
CCL-134HumanIPF pulmonary fibroblast
CCL-190HumanPulmonary fibroblast
FU-UR-1HumanRenal cell carcinoma
H9c2RatCardiac myoblast
HCLEHumanCorneal epithelial
HeLaHumanCervical cancer
HMECHumanMicrovascular endothelial
HT-1080HumanBreast fibrosarcoma
JYHumanLymphoblastic leukemia
K562HumanMyelogenous leukaemia
LLC-PK1PigKidney epithelial
MCF-7HumanBreast adenocarcinoma
MCF-10AHumanBreast adenocarcinoma
MDA-MB-231HumanBreast adenocarcinoma
MDA-MB-468HumanBreast cancer
MDCKCanineKidney epithelial
MES-SAHumanUterine sarcoma
MES-SA/DXHumanUterine sarcoma
Min 6MousePancreatic insulinoma
NRKRatKidney epithelial
NRK-52ERatKidney epithelial
PC-3HumanProstate cancer
SKBR3HumanBreast cancer
STHdhMouseStriatal cells
TK6HumanSplenic lymphoblast
U373 MGHumanGlioblastoma
WEHI 7.2MouseLymphoid

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The substrates are very stable. Some users have reported performing time course assays with NucView® 488 Caspase-3 Substrate for 4-5 days.

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