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NUCVIEW® CASPASE-3 SUBSTRATES

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

HOW IT WORKS

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.

Features

  • 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 cell 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).

THE GOLD STANDARD FOR MONITORING APOPTOSIS

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. doi.org/10.1016/j.ccell.2016.10.010

DUAL DETECTION KITS AVAILABLE

Planning on using NucView® Caspase-3 Substrates with other markers?

See our dual detection apoptosis kits which pair NucView® 488 with either Annexin V conjugates, MitoView™ 633, or RedDot™2.

  • Dual Apoptosis Kit: NucView® 488 and Annexin V labeled with red or far-red dyes for co-detection of two apoptotic events, caspase cleavage and phosphatidylserine (PS) translocation.
  • Dual Apoptosis Kit: NucView® 488 and MitoView™ 633 for co-detection of two apoptotic events, caspase cleavage and loss of mitochondrial membrane potential.
  • Apoptosis/Necrosis Kit: NucView® 488 and RedDot™2 for concurrent measurement of caspase cleavage (apoptosis) and loss of membrane integrity (necrosis).

MCF-7 cells were left untreated or treated with staurosporine to induce apoptosis, then with 40 nM MitoView™ Blue, NucView® 520 Caspase-3 Substrate, and CF® 488A Annexin V. Healthy cells show mitochondrial staining with MitoView™ Blue (cyan), while apoptotic cells show NucView® 520 staining of nuclei (red) and CF® 488A Annexin V staining of cell membranes (green).

NucView® Caspase-3 Substrates and Kits

Catalog No.

Features

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

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

From: $362 Sizes: 50 assaysCatalog #: 30030, 30067, - 30073View allHide

NucView® 405 Caspase-3 Substrate, 1 mM in DMSO

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

NucView® 488 & MitoView™ 633 Apoptosis Assay Kit

From: $351 Sizes: 100 assaysCatalog #: 30062

NucView® 488 & RedDot™ 2 Apoptosis & Necrosis Kit

From: $308 Sizes: 100 assaysCatalog #: 30072

NucView® 488 Caspase-3 Assay Kit for Live Cells

From: $98 Sizes: Trial size (25 assays), 100 assaysCatalog #: 30029, - 30029-TView allHide

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

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

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

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

NucView® 530 Caspase-3 Substrate, 1 mM in DMSO

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

 

Faq

NucView® Caspase-3 Enzyme Substrates

The substrates are 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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