Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) ab219915 is used to monitor multiple caspase activation in live cells
- 3 fluorogenic indicators for simultaneous monitoring of caspase3/7, caspase 8 and caspase 9
- Optimised for microplate reader
- Up to 100 tests for each caspase
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- ChemMedChem 19:e2023006802024Effective Anticancer Potential of a New Sulfonamide as a Carbonic Anhydrase IX Inhibitor Against Aggressive Tumors.Applications:Unspecified applicationReactive species:Unspecified reactive speciesIsmail Koyuncu,Ebru Temiz,Eray Metin Güler,Mustafa Durgun,Ozgür Yuksekdag,Simone Giovannuzzi,Claudiu T SupuranPubMed 38323458
- International journal of nanomedicine 18:4253-42742023A Dual Concentration-Tailored Cytokine-Chemo Nanosystem to Alleviate Multidrug Resistance and Redirect Balance of Cancer Proliferation and Apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYu Hsia,Maharajan Sivasubramanian,Chia-Hui Chu,Yao-Chen Chuang,Yiu-Kay Lai,Leu-Wei LoPubMed 37534057
- Toxins 15:2023Engineering BinB Pore-Forming Toxin for Selective Killing of Breast Cancer Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTipaporn Kumkoon,Chalongrat Noree,Panadda BoonsermPubMed 37104235
- Life sciences 315:1213582023Oxidative stress stimulation leads to cell-specific oxidant and antioxidant responses in airway resident and inflammatory cells.Applications:Unspecified applicationReactive species:HumanHayriye Akel Bilgic,Busra Kilic,Berfin Doga Kockaya,Basak Ezgi Sarac,Aysun Kilic Suloglu,Omer Kalayci,Cagatay KaraaslanPubMed 36596408
- Cells 11:2022In Vitro Cell Death Mechanisms Induced by Root Extract in Combination with ZnPcS Mediated-Photodynamic Therapy in A549 Lung Cancer Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlexander Chota,Blassan P George,Heidi AbrahamsePubMed 36291155
- Frontiers in oncology 12:9585362022Distinct mechanism of cervical cancer cell death caused by the investigational new drug SHetA2.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRajani Rai,Vishal Chandra,Amy L Kennedy,Rosemary E Zuna,Doris Mangiaracina BenbrookPubMed 36203464
- Communications biology 5:7162022Cytoplasmic sequestration of p53 by lncRNA-CIRPILalleviates myocardial ischemia/reperfusion injury.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYuan Jiang,Ying Yang,Yang Zhang,Jiqin Yang,Man-Man Zhang,Shangxuan Li,Genlong Xue,Xingda Li,Xiaofang Zhang,Jiming Yang,Xiang Huang,Qihe Huang,Hongli Shan,Yanjie Lu,Baofeng Yang,Zhenwei PanPubMed 35851102
- Clinical and translational medicine 12:e9312022ISG15 deficiency features a complex cellular phenotype that responds to treatment with itaconate and derivatives.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSyed Fakhar-Ul-Hassnain Waqas,Aaqib Sohail,Ariane Hai Ha Nguyen,Abdulai Usman,Tobias Ludwig,Andre Wegner,Muhammad Nasir Hayat Malik,Sven Schuchardt,Robert Geffers,Moritz Winterhoff,Sylvia Merkert,Ulrich Martin,Ruth Olmer,Nico Lachmann,Frank PesslerPubMed 35842904
- Designed monomers and polymers 25:197-2042022Two new Cd(II)/Zn(II) coordination polymers: luminescence properties and synergistic treatment activity with ultrasound therapy on uterine fibroids.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHong-Mei Liu,Xiao-Na ShangPubMed 35755879
- Translational cancer research 10:424-4332022Determining the effect of ellagic acid on the proliferation and migration of pancreatic cancer cell lines.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJi Yoon Kim,Yun Ju Choi,Ha-Jeong KimPubMed 35116272
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Semi-quantitative
Target data
Function
Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:35338844, PubMed:35446120, PubMed:7596430). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:7596430). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond (PubMed:10497198, PubMed:16374543, PubMed:7596430, PubMed:7774019). Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain (By similarity). Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively) (PubMed:7596430). Cleaves and inactivates interleukin-18 (IL18) (PubMed:37993714, PubMed:9334240). Involved in the cleavage of huntingtin (PubMed:8696339). Triggers cell adhesion in sympathetic neurons through RET cleavage (PubMed:21357690). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:23152800). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (PubMed:35338844, PubMed:35446120). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:23845944, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758104, PubMed:36758106).
Additional Targets
CASP8, CASP9, CASP7
Alternative names
CPP32, CASP3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1
What's included?
Recommended products
Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) ab219915 is used to monitor multiple caspase activation in live cells
- 3 fluorogenic indicators for simultaneous monitoring of caspase3/7, caspase 8 and caspase 9
- Optimised for microplate reader
- Up to 100 tests for each caspase
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Semi-quantitative
- Assay Platform
- Microplate reader
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) is designed to simultaneously monitor three key caspases involved in apoptosis: the initiator caspases caspase 8 and caspase 9, and the executioner Caspase 3.
How the assay works
The kit uses DEVD-ProRed™, IETD-R110 and LEHD-AMC as fluorogenic indicators for Caspase 3, caspase 8 and caspase 9 activity respectively. Upon caspase cleavage, three distinct fluorophores are released: ProRed™ (red fluorescence), R110 (green fluorescence) and AMC (blue fluorescence), which can be readily monitored in a single assay due to their spectral separation.
This product has been optimized for use in a microplate reader in a 96-well plate, providing enough reagent to perform 100 tests for each caspase.
Caspase-3/7, Caspase-8 and Caspase-9 protocol summary
- Grow and treat cells with test compounds to induce caspase activation
- Add caspase assay solution to cells
- Incubate at room temperature for 30-60 minutes
- Monitor fluorescence intensity at: Ex/Em = 535/620 nm (Caspase 3/7), Ex/Em = 490/525 nm (Caspase 8), Ex/Em = 370/450 nm (Caspase 9)
How other researchers are using this kit
ab219915 has been used in a variety of cancer cell lines including MCF-7 (breast cancer) cell line (1), U87MG (human glioblastoma) and MES-SA (human uterine sarcoma) cell lines (2).
References: 1 - T. Kumkoon et al. 2023; 2 - Yu Hsia et al. 2023.
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3 product images
Functional Studies - Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) (ab219915)
Detection of Caspase activity in Jurkat cells Caspase 3.
Caspase 3, Caspase 8 and Caspase 9 Multiplex Activity Assay Kit (Fluorometric) (ab219915). Detection of Caspase activity in Jurkat cells graphs shown Caspase 3 activity. Jurkat cells were seeded on the same day at 2 x 105 cells/well in a Costar black wall/clear bottom 96-well plate. Cells were either left untreated (black bar) or treated with 1 μM staurosporine for 4 hours (gray bar). Single-caspase assay loading solution (100 μL/well; #1 for Caspase 3, #2 for caspase 8 or #3 for caspase 9) or Triple-caspase assay loading solution (100 μL/well; #4 for Caspase 3, 8 and 9 together) was added to cells, followed by an incubation at RT for 1 hour. The fluorescence intensity was measured with FlexStation fluorescence microplate reader at the indicated wavelength. Caspase 3, 8 and 9 activities can be detected in a single assay without interferences from other caspases.
Functional Studies - Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) (ab219915)
Detection of Caspase activity in Jurkat cells Caspase 9.
Caspase 3, Caspase 8 and Caspase 9 Multiplex Activity Assay Kit (Fluorometric) (ab219915). Detection of Caspase activity in Jurkat cells graphs shown Caspase 9 activity. Jurkat cells were seeded on the same day at 2 x 105 cells/well in a Costar black wall/clear bottom 96-well plate. Cells were either left untreated (black bar) or treated with 1 μM staurosporine for 4 hours (gray bar). Single-caspase assay loading solution (100 μL/well; #1 for Caspase 3, #2 for caspase 8 or #3 for caspase 9) or Triple-caspase assay loading solution (100 μL/well; #4 for Caspase 3, 8 and 9 together) was added to cells, followed by an incubation at RT for 1 hour. The fluorescence intensity was measured with FlexStation fluorescence microplate reader at the indicated wavelength. Caspase 3, 8 and 9 activities can be detected in a single assay without interferences from other caspases.
Functional Studies - Caspase-3/7, Caspase-8 and Caspase-9 Multiplex Activity Assay Kit (Fluorometric) (ab219915)
Detection of Caspase activity in Jurkat cells Caspase 8.
Caspase 3, Caspase 8 and Caspase 9 Multiplex Activity Assay Kit (Fluorometric) (ab219915). Detection of Caspase activity in Jurkat cells graphs shown Caspase 8 activity. Jurkat cells were seeded on the same day at 2 x 105 cells/well in a Costar black wall/clear bottom 96-well plate. Cells were either left untreated (black bar) or treated with 1 μM staurosporine for 4 hours (gray bar). Single-caspase assay loading solution (100 μL/well; #1 for Caspase 3, #2 for caspase 8 or #3 for caspase 9) or Triple-caspase assay loading solution (100 μL/well; #4 for Caspase 3, 8 and 9 together) was added to cells, followed by an incubation at RT for 1 hour. The fluorescence intensity was measured with FlexStation fluorescence microplate reader at the indicated wavelength. Caspase 3, 8 and 9 activities can be detected in a single assay without interferences from other caspases.
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