Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Blue Fluorescence*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Excitation (nm) | 341 |
Emission (nm) | 441 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Alternative formats
Related products
Overview | ![]() ![]() |
See also: Caspases
Excitation (nm) 341 | Emission (nm) 441 |
Our Cell Meter™ assay kits are a set of tools for monitoring cell viability. There are a variety of parameters that can be used for monitoring cell viability. This particular kit is designed to monitor cell apoptosis through measuring Caspase 3 activation. Caspase 3 is widely accepted as a reliable indicator for cell apoptosis since the activation of caspase-3 (CPP32/apopain) is important for the initiation of apoptosis. Caspase 3 has substrate selectivity for the peptide sequence Asp-Glu-Val-Asp (DEVD). This kit uses Ac-DEVD-AMC as a fluorogenic indicator for caspase-3 activity. Cleavage of AMC peptides by caspase 3 generates strongly fluorescent AMC that is monitored fluorimetrically at 450-480 nm with excitation of 340-370 nm. The kit provides all the essential components with an optimized assay protocol. The assay is robust, and can be readily adapted for high-throughput assays. Using 100 uL of reagents per well in a 96-well format, this kit provides sufficient reagents to perform 200 assays. Using 25 uL of reagents per well in a 384-well format, this kit provides sufficient reagents to perform 800 assays.
Platform
Fluorescence microplate reader
Excitation | 360 nm |
Emission | 470 nm |
Cutoff | 420 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Top/Bottom read mode |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds (100 µL/well/96-well plate or 25 µL/well/384-well plate)
- Add equal volume of Caspase 3/7 Substrate working solution
- Incubate at room temperature for 1 hour
- Monitor fluorescence intensity at Ex/Em = 350/450 nm (Cutoff = 420 nm)
Important notes
Thaw one vial of each kit component at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 50 µL of Caspase 3/7 Substrate (Component A) into 10 mL of Assay Buffer (Component B) and mix well to make Caspase 3/7 Substrate working solution. Note: Aliquot and store the unused Components A and B at -20 oC. Avoid repeated freeze/thaw cycles.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells by adding 10 µL/well of 10X test compounds (96-well plate) or 5 µL/well of 5X test compounds (384-well plate) into PBS or the desired buffer. For blank wells (medium without the cells), add the same amount of compound buffer.
- Incubate the cell plate in a 37°C, 5% CO2 incubator for a desired period of time (4 - 6 hours for Jurkat cells treated with camptothecin) to induce apoptosis.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Caspase 3/7 Substrate working solution.
- Incubate the plate at room temperature for at least 1 hour, protected from light. Note: If desired, add 1 µL of the 1 mM Ac-DEVD-CHO caspase 3/7 inhibitor into selected samples 10 minutes before adding Caspase 3/7 working solution at room temperature to confirm the inhibition of the caspase 3/7-like activities.
- Centrifuge cell plate (especially for the non-adherent cells) at 800 rpm for 2 minutes (brake off).
- Monitor the fluorescence intensity with a fluorescence microplate reader at Ex/Em = 350/450 nm (Cutoff = 420 nm).
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) |
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence* | 500 | 522 | 80000 |
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Red Fluorescence* | 532 | 619 | - |
Images

Figure 1. Detection of Caspase 3/7 activity in Jurkat cells with Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit. Jurkat cells were seeded on the same day at 80,000 cells/well/90 µL in a Costar black wall/clear bottom 96-well plate. The cells were treated with or without 1 µM of staurosporine for 4 hours, and with or without 10 µM of the caspase inhibitor AC-DEVD-CHO for 10 minutes. The caspase 3/7 assay solution (100 µL/well) was added and incubated at room temperature for 1 hour. The fluorescence intensity was measured at Ex/Em = 360/470 nm (Cutoff = 420 nm).

Figure 2. Albumin exposure upregulates pro-apoptotic pathways in podocytes. A, Activated caspase 3/7 activity normalized to total cellular protein in podocytes treated with 5 mg/ml albumin (closed bars) or 5 mg/ml dextran (open bars) as an oncotic control for the indicated amounts of time. * denotes P<0.0001 compared to corresponding dextran treated control. B, TUNEL staining of cultured podocytes treated with 5 mg/ml dextran (Dex) or 5 mg/ml albumin (Alb). Nuclei are stained blue with DAPI. TUNEL-positive nuclei are green. C, Caspase 3/7 activity normalized to total cellular protein in podocytes treated with 5 mg/ml albumin or 5 mg/ml albumin +50 µM z-VAD, a pan-caspase inhibitor. z-VAD largely abrogated caspase activity (*, P = 0.01 versus albumin+z-VAD). D, Percentage of dead cells measured using the trypan blue assay in podocytes treated with 5 mg/ml albumin alone (open bar) or 5 mg/ml albumin +50 µM z-VAD (closed bar) for 24 hrs or 5 mg/ml albumin alone (horizontal hatched bar) or 5 mg/ml albumin +50 µM z-VAD for 48 hrs (vertical hatched bar). * denotes P = 0.001 compared to albumin+z-VAD at 48 hrs. *Caspase activity was determined using the Caspase 3/7 Activity kit (AAT Bioquest, Sunnyvale, CA) according to the manufacturer’s directions. Source: Graph from Endocytosis of Albumin by Podocytes Elicits an Inflammatory Response and Induces Apoptotic Cell Death by Kayo Okamura, et al., PLoS ONE, Jan. 2013.
Citations
View all 36 citations: Citation Explorer
Trans-[bis (benzimidazol-2-ylidene) dichlorido] platinum (II) complexes with peculiar modes of action and activity against cisplatin-resistant cancer cells
Authors: B{\"a}r, Sofia I and Schleser, Sebastian W and Oberhuber, Natalie and Herrmann, Alexander and Schlotte, Luca and Weber, Stefanie E and Schobert, Rainer
Journal: Journal of Inorganic Biochemistry (2023): 112028
Authors: B{\"a}r, Sofia I and Schleser, Sebastian W and Oberhuber, Natalie and Herrmann, Alexander and Schlotte, Luca and Weber, Stefanie E and Schobert, Rainer
Journal: Journal of Inorganic Biochemistry (2023): 112028
Involvement of necroptosis in the selective toxicity of the natural compound ($\pm$) gossypol on squamous skin cancer cells in vitro
Authors: Haasler, Lisa and von Montfort, Claudia and Kondadi, Arun Kumar and Golombek, Mathias and Ebbert, Lara and Wenzel, Chantal-Kristin and Stahl, Wilhelm and Reichert, Andreas S and Brenneisen, Peter
Journal: Archives of Toxicology (2023): 1--18
Authors: Haasler, Lisa and von Montfort, Claudia and Kondadi, Arun Kumar and Golombek, Mathias and Ebbert, Lara and Wenzel, Chantal-Kristin and Stahl, Wilhelm and Reichert, Andreas S and Brenneisen, Peter
Journal: Archives of Toxicology (2023): 1--18
Metformin sensitizes AML cells to venetoclax through endoplasmic reticulum stress—CHOP pathway
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Apical-Out Chicken Enteroids as an In-Vitro Model to Study Effects of an Alternative Growth Promoter on Indicators of Gut Health
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Journal: (2023)
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Journal: Journal of Health and Translational Medicine (JUMMEC) (2023): 105--115
Authors: Bakar, Siti Aishah Abu and Azhar, Nur Asna and Mohamad, Noor Muzamil and Nordin, Ira Maya Sophia and Ali, Abdul Manaf and Noor, Siti Noor Fazliah Mohd and Hamid, Shahrul Bariyah Sahul and Ahmad, Nor Hazwani
Journal: Journal of Health and Translational Medicine (JUMMEC) (2023): 105--115
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Journal: Scientific reports (2022): 1--10
Authors: Jo, Ahyoung and Kwak, Jae-Hwan and Woo, Soo-Yeon and Kim, Bo-Young and Son, Yonghae and Choi, Hee-Seon and Kim, Jayoung and Kwon, Munju and Cho, Hyok-Rae and Eo, Seong-Kug and others,
Journal: Scientific reports (2022): 1--10
Revisiting the anticancer properties of phosphane (9-ribosylpurine-6-thiolato) gold (I) complexes and their 9H-purine precursors
Authors: Kober, Luisa and Schleser, Sebastian W and B{\"a}r, Sofia I and Schobert, Rainer
Journal: JBIC Journal of Biological Inorganic Chemistry (2022): 731--745
Authors: Kober, Luisa and Schleser, Sebastian W and B{\"a}r, Sofia I and Schobert, Rainer
Journal: JBIC Journal of Biological Inorganic Chemistry (2022): 731--745
Visfatin inhibits colon cancer cell apoptosis and decreases chemosensitivity to 5-FU by promoting the SDF-1/CXCR4/Akt axis
Authors: Zhao, Quan and Long, Yaxin and Cheng, Wen and Huang, Yingguang and Li, Jinyuan and Li, Yuejin and Li, Xing and Guo, Xiaodong and Li, Yu and Li, Guosan and others,
Journal: International journal of oncology (2022): 1--13
Authors: Zhao, Quan and Long, Yaxin and Cheng, Wen and Huang, Yingguang and Li, Jinyuan and Li, Yuejin and Li, Xing and Guo, Xiaodong and Li, Yu and Li, Guosan and others,
Journal: International journal of oncology (2022): 1--13
Melatonin alleviates alcoholic liver disease via EGFR--BRG1--TERT axis regulation
Authors: Che, Zhaodi and Song, Yali and Xu, Chengfang and Li, Wei and Dong, Zhiyong and Wang, Cunchuan and Ren, Yixing and So, Kwok-Fai and Tipoe, George L and Wang, Fei and others,
Journal: Acta Pharmaceutica Sinica B (2022)
Authors: Che, Zhaodi and Song, Yali and Xu, Chengfang and Li, Wei and Dong, Zhiyong and Wang, Cunchuan and Ren, Yixing and So, Kwok-Fai and Tipoe, George L and Wang, Fei and others,
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Combination of Goniothalamin and Sol-Gel-Derived Bioactive Glass 45S5 Enhances Growth Inhibitory Activity via Apoptosis Induction and Cell Cycle Arrest in Breast Cancer Cells MCF-7
Authors: Bakar, Siti Aishah Abu and Ali, Abdul Manaf and Noor, Siti Noor Fazliah Mohd and Hamid, Shahrul Bariyah Sahul and Azhar, Nur Asna and Mohamad, Noor Muzamil and Ahmad, Nor Hazwani
Journal: BioMed Research International (2022)
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Journal: BioMed Research International (2022)
Application notes
FAQ
Are inflammasomes and caspase-1 related?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?
Do you offer any fluorimetric assays that measure caspase activation/activity in live cells using a flow cytometer?
Does pH and staining temperature affect Annexin V-Phosphatidylserine binding?
Does propidium iodide stain apoptotic cells?
How can I tell if my cell sample is dying?