Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
Quotation | Request |
International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Extinction coefficient (cm -1 M -1) | 80000 |
Excitation (nm) | 500 |
Emission (nm) | 522 |
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
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Overview | ![]() ![]() |
Extinction coefficient (cm -1 M -1) 80000 | Excitation (nm) 500 | Emission (nm) 522 |
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 Z-DEVD-Rh 110-DVED-Z as a fluorogenic indicator for caspase-3 activity. Cleavage of Rh 110 peptides by caspase 3 generates strongly fluorescent Rh 110 that is monitored fluorimetrically at 520-530 nm with excitation of 480-500 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 | 490 nm |
Emission | 525 nm |
Cutoff | 515 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 = 490/525 nm (Cutoff = 515 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 Caspase 3/7 Substrate (Components A) and Assay Buffer (Component B) at -20 oC. Avoid repeated freeze/thaw cycles
SAMPLE EXPERIMENTAL PROTOCOL
Cells preparation:
- For adherent cells: Plate cells overnight in growth medium at 20,000 cells/well/90 uL for a 96-well plate or 5,000cells/well/20mL for a 384-well plate.
- For non-adherent cells: Centrifuge the cells from the culture medium and then suspend the cell pellet in culture medium at 80,000 cells/well/90 uL for a 96-well poly-D lysine plate or 20,000 cells/well/20 uL for a 384-well poly-D lysine plate. Centrifuge the plate at 800 rpm for 2 minutes with brake off prior to the experiments. Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.
Sample 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 = 490/525 nm (Cutoff = 515 nm).
Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Extinction coefficient (cm -1 M -1) | 80000 |
Excitation (nm) | 500 |
Emission (nm) | 522 |
Product Family
Name | Excitation (nm) | Emission (nm) |
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Blue Fluorescence* | 341 | 441 |
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. Jurkat cells were seeded on the same day at 80,000 cells/well/90 µL in a black wall/clear bottom 96-well costar plate. The cells were treated with or without 20 µM of camptothecin for 5 hours, and/or 5 µM caspase 3/7 inhibitor AC-DEVD-CHO for 10 minutes. The caspase 3/7 working solution (100 µL/well) was added and incubated at room temperature for 1 hour. The fluorescence intensity was measured at Ex/Em = 490/525 nm (Cutoff = 515 nm) using the NOVOstar instrument (BMG Labtech).
Citations
View all 31 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
Oxime derivative TFOBO promotes cell death by modulating reactive oxygen species and regulating NADPH oxidase activity in myeloid leukemia
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
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
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)
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)
SPR064, a pro-drug of paclitaxel, has anti-tumorigenic effects in endometrial cancer cell lines and mouse models
Authors: Zhao, Xiaoling and Kong, Weimin and Tucker, Katherine and Staley, Allison and Fan, Yali and Sun, Wenchuan and Yin, Yajie and Huang, Yu and Fang, Ziwei and Wang, Jiandong and others,
Journal: American Journal of Translational Research (2020): 4264
Authors: Zhao, Xiaoling and Kong, Weimin and Tucker, Katherine and Staley, Allison and Fan, Yali and Sun, Wenchuan and Yin, Yajie and Huang, Yu and Fang, Ziwei and Wang, Jiandong and others,
Journal: American Journal of Translational Research (2020): 4264
Dendrimer Conjugation Enhances Tumor Penetration and Efficacy of Doxorubicin in Extracellular Matrix-Expressing 3D Lung Cancer Models
Authors: Almuqbil, Rashed M and Heyder, Rodrigo S and Bielski, Elizabeth R and Durymanov, Mikhail and Reineke, Joshua J and da Rocha, Sandro RP
Journal: Molecular Pharmaceutics (2020): 1648--1662
Authors: Almuqbil, Rashed M and Heyder, Rodrigo S and Bielski, Elizabeth R and Durymanov, Mikhail and Reineke, Joshua J and da Rocha, Sandro RP
Journal: Molecular Pharmaceutics (2020): 1648--1662
microRNA-144 inhibits cell proliferation and invasion by directly targeting TIGAR in esophageal carcinoma
Authors: Mu, Yushu and Wang, Qifei and Tan, Lei and Lin, Lin and Zhang, Benhua
Journal: Oncology letters (2020): 3079--3088
Authors: Mu, Yushu and Wang, Qifei and Tan, Lei and Lin, Lin and Zhang, Benhua
Journal: Oncology letters (2020): 3079--3088
The potential role of extracellular vesicles in COVID-19 associated endothelial injury and pro-inflammation
Authors: Krishnamachary, Balaji and Cook, Christine and Spikes, Leslie and Chalise, Prabhakar and Dhillon, Navneet K
Journal: medRxiv (2020)
Authors: Krishnamachary, Balaji and Cook, Christine and Spikes, Leslie and Chalise, Prabhakar and Dhillon, Navneet K
Journal: medRxiv (2020)
Therapeutic potential of KLF2-induced exosomal microRNAs in pulmonary hypertension
Authors: Sindi, Hebah A and Russomanno, Giusy and Satta, Sandro and Abdul-Salam, Vahitha B and Jo, Kyeong Beom and Qazi-Chaudhry, Basma and Ainscough, Alexander J and Szulcek, Robert and Bogaard, Harm Jan and Morgan, Claire C and others,
Journal: Nature communications (2020): 1--17
Authors: Sindi, Hebah A and Russomanno, Giusy and Satta, Sandro and Abdul-Salam, Vahitha B and Jo, Kyeong Beom and Qazi-Chaudhry, Basma and Ainscough, Alexander J and Szulcek, Robert and Bogaard, Harm Jan and Morgan, Claire C and others,
Journal: Nature communications (2020): 1--17
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?
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