Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Blue Fluorescence*
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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 |
Excitation (nm) | 341 |
Emission (nm) | 441 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Excitation (nm) 341 | Emission (nm) 441 |
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
Prepare cells with test compounds (100 µL/well/96-well plate or 25 µL/well/384-well plate).
Add an equal volume of Caspase 3/7 Substrate working solution.
Incubate at room temperature for 1 hour.
Monitor fluorescence intensity at Ex/Em = 360/470 nm (Cutoff = 420 nm).
Thaw one vial of each kit component at room temperature before starting the experiment.
CELL PREPARATION
For guidelines on cell sample preparation, please visit:
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
PREPARATION OF WORKING SOLUTION
Prepare a Caspase 3/7 Substrate working solution by combining 50 µL of Caspase 3/7 Substrate (Component A) with 10 mL of Assay Buffer (Component B), and mix well.
Note: Aliquot and store any unused Components A and B at -20°C. Avoid repeated freeze/thaw cycles.
SAMPLE EXPERIMENTAL PROTOCOL
To treat cells, add 10 µL per well of 10X test compounds for a 96-well plate or 5 µL per well of 5X test compounds for a 384-well plate into PBS or the desired buffer. For blank wells containing only medium, add an equivalent volume of the compound buffer.
Incubate the cell plate at 37°C with 5% CO2 for a desired duration to induce apoptosis (e.g., 4 to 6 hours for Jurkat cells treated with camptothecin). Adjust the time as necessary based on the cell type and treatment conditions.
Add 100 µL/well for a 96-well plate or 25 µL/well for a 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: To confirm the inhibition of the caspase 3/7-like activities, add 1 µL of 1 mM Ac-DEVD-CHO caspase 3/7 inhibitor to selected samples 10 minutes before adding the Caspase 3/7 working solution at room temperature.
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 = 360/470 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
![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).](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-caspase-3-7-activity-apoptosis-assay-kit-blue-fluorescence%2Ffigure-for-cell-meter-caspase-3-7-activity-apoptosis-assay-kit-blue-fluorescence_UJkSj.jpg&w=3840&q=75)
![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 <strong>Endocytosis of Albumin by Podocytes Elicits an Inflammatory Response and Induces Apoptotic Cell Death</strong> by Kayo Okamura, et al., <em>PLoS ONE</em>, Jan. 2013.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-meter-caspase-3-7-activity-apoptosis-assay-kit-blue-fluorescence%2Ffigure-for-cell-meter-caspase-3-7-activity-apoptosis-assay-kit-blue-fluorescence_70uWq.jpg&w=3840&q=75)
Citations
Authors: Shen, Xiaochang and Wang, Jiandong and Deng, Boer and Chen, Shuning and John, Catherine and Zhao, Ziyi and Sinha, Nikita and Haag, Jennifer and Sun, Wenchuan and Kong, Weimin and others,
Journal: Gynecologic Oncology (2024): 93--102
Authors: Jiang, Xinya and Huang, Kexiu and Sun, Xiaofan and Li, Yue and Hua, Lei and Liu, Fangshu and Huang, Rui and Du, Juan and Zeng, Hui
Journal: iScience (2023)
Authors: Tamazian, Lorik
Journal: (2023)
Authors: Hua, Lei and Yang, Nianhui and Li, Yue and Huang, Kexiu and Jiang, Xinya and Liu, Fangshu and Yu, Zhi and Chen, Jie and Lai, Jing and Du, Juan and others,
Journal: British Journal of Haematology (2023)
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
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: 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: 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: 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: 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
Application notes
FAQ
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?