Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

Cell Meter™ No-Wash Live Cell Caspase 3/7 Activity Assay Kit *Red Fluorescence*

The fluorescence microscope images of normal Hela cells (A) and apoptotic Hela cells (B). Hela cells were cultured in a 96-well plate, and washed twice with HHBS buffer.  ApoBrite™ V570 caspase 3/7 dye loading solution was then added to the well. After incubation for 2 h at 37 °C, the cells were washed once with HHBS buffer and treated with staurosporine (1 μM) apoptosis inducer for 1 hr. The images were acquired using a fluorescence microscope equipped with DAPI filter set.
The fluorescence microscope images of normal Hela cells (A) and apoptotic Hela cells (B). Hela cells were cultured in a 96-well plate, and washed twice with HHBS buffer.  ApoBrite™ V570 caspase 3/7 dye loading solution was then added to the well. After incubation for 2 h at 37 °C, the cells were washed once with HHBS buffer and treated with staurosporine (1 μM) apoptosis inducer for 1 hr. The images were acquired using a fluorescence microscope equipped with DAPI filter set.
Ordering information
Price ()Discontinued
Catalog Number20260
Unit Size
Find Distributor
Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Related products
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Green Fluorescence*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Blue Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Intracellular Fluorimetric Hydrogen Peroxide Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrial Hydroxyl Radical Detection Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit *Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Orange Fluorescence Optimized for Microplate Reader*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Live Cell Caspase 3/7 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 3/7 Binding Assay Kit *Red Fluorescence*
Cell Meter™ Live Cell Caspase 1 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 2 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 6 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 8 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 9 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 10 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Live Cell Caspase 13 Binding Assay Kit *Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular pH Assay Kit
Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit
Cell Meter™ Cell Viability Assay Kit *Red Fluorescence*
Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence with 405 nm Excitation*
Cell Meter™ Cell Viability Assay Kit *Blue Fluorescence*
Cell Meter™ Cell Viability Assay Kit *Green Fluorescence*
Cell Meter™ Cell Viability Assay Kit *NIR Fluorescence Optimized for Fluorescence Microplate Reader*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Blue Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Red Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Microplate Readers*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Orange Fluorescence Optimized for Microplate Readers*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Microplate Assays*
Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry Assays*
Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ NIR Mitochondrion Membrane Potential Assay Kit *Optimized for Microplate Reader*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Orange Fluorescence Optimized for Microplate Reader*
Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Nuclear Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Multiplexing Caspase 3/7, 8 and 9 Activity Assay Kit *Triple Fluorescence Colors*
Cell Meter™ Generic Fluorimetric Caspase Activity Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Generic Fluorimetric Caspase Binding Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Orange Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Blue Fluorescence Excited at 405 nm*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Green Fluorescence Excited at 405 nm*
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Orange Fluorescence Excited at 405 nm*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Blue Fluorescence Excited at 405 nm*
Cell Meter™ Phosphatidylserine Apoptosis Assay Kit *Green Fluorescence Excited at 405 nm*
Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ PE-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Apoptotic and Necrotic Multiplexing Detection Kit I *Triple Fluorescence Colors*
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Fixed Cell Cycle Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Apoptotic and Necrotic Multiplexing Detection Kit II *Triple Fluorescence Colors*
Cell Meter™ Live Cell TUNEL Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Optimized for 405 nm Violet Laser Excitation*
Cell Meter™ Live Cell Caspase 3/7 and Phosphatidylserine Detection Kit *Triple Fluorescence Colors*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Green Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Orange Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Deep Red Fluorescence*
Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit*Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Mitochondrial Superoxide Activity Assay Kit*Optimized for Microplate Reader*
Cell Meter™ Autophagy Assay Kit *Blue Fluorescence*
Cell Meter™ Autophagy Fluorescence Imaging Kit
Cell Meter™ Autophagy Assay Kit *Green Fluorescence*
Cell Meter™ FITC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *NIR Fluorescence Optimized for Microplate Reader*
Cell Meter™ Fluorimetric Intracellular Nitric Oxide (NO) Activity Assay Kit *NIR Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Colorimetric WST-8 Cell Quantification Kit
Cell Meter™ Multiplexing Live, Apoptotic and Necrotic Cell Detection Kit III *Triple Fluorescence Colors*
Cell Meter™ 2-NBDG Glucose Uptake Assay Kit
Cell Meter™ Intracellular NADH/NADPH Fluorescence Imaging Kit *Red Fluorescence*
Cell Meter™ Intracellular NADH/NADPH Flow Cytometric Analysis Kit *Red Fluorescence*
Cell Meter™ Live Cell TUNEL Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit *Optimized for microplate reader*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Mitochondrion Membrane Potential Assay Kit *Red Fluorescence Optimized for Microplate Reader*
Cell Meter™ Intracellular Colorimetric Lipid Peroxidation (MDA) Assay Kit
Cell Meter™ Flow Cytometric Calcium Assay Kit
Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Red Fluorescence Optimized for Flow Cytometry*
Cell Meter™ Fluorimetric Cellular Lipid Peroxidation Assay Kit
Cell Meter™ Intracellular NADH/NADPH Fluorescence Imaging Kit *Deep Red Fluorescence*
Cell Meter™ Intracellular NADH/NADPH Flow Cytometric Analysis Kit *Deep Red Fluorescence*
Cell Meter™ Intracellular GSH Assay Kit *Optimized for Flow Cytometry with 405 nm excitation*
Cell Meter™ Mitochondrial Autophagy Imaging Kit *Red Fluorescence*
Cell Meter™ BX520 fixable viability dye
Cell Meter™ BX590 fixable viability dye
Cell Meter™ BX650 fixable viability dye
Cell Meter™ IX830 fixable viability dye
Cell Meter™ RX660 fixable viability dye
Cell Meter™ RX700 fixable viability dye
Cell Meter™ RX780 fixable viability dye
Cell Meter™ VX450 fixable viability dye
Cell Meter™ VX500 fixable viability dye
Cell Meter™ Live Cell Caspase 8 Binding Assay Kit *Red Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Green Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Red Fluorescence*
Cell Meter™ Fluorimetric Cellular Voltage Assay Kit
Cell Meter™ Cell Adhesion Assay Kit
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Deep Red Fluorescence*
Cell Meter™ Fixed Cell and Tissue TUNEL Apoptosis Assay Kit *Blue Fluorescence*
Cell Meter™ Fluorescence Gap Junction Tracing Kit
Cell Meter™ Colorimetric MTT Cell Proliferation Kit
Cell Meter™ Live Cell Caspase 3/7 Imaging Kit *Green Fluorescence*
Cell Meter™ Beta-Arrestin Translocation GPCR Signaling Kit
Cell Meter™ Live Cell ATP Assay Kit
Cell Meter™ Colorimetric Cell Cytotoxicity Assay Kit
Cell Meter™ Fluorimetric Phagocytosis Assay Kit *Red Fluorescence*
Cell Meter™ Cellular Senescence Activity Assay Kit *Green Fluorescence*
Cell Meter™ Cellular Senescence Activity Assay Kit *Red Fluorescence*
Cell Meter™ Cell Proliferation Assay Kit
Show More (129)

OverviewpdfSDSpdfProtocol


The activation of caspase 3/7 is important for the initiation of apoptosis. Our Cell Meter™ No-Wash Live Cell Caspase Activity Assay Kits are based on ApoBrite™ V600, our recently developed cell-permeable fluorogenic caspase substrate, the first fluorogenic probe for the direct detection of caspase activities in live cells. ApoBrite V600 consists of three moieties including a). masked fluorophore, b). caspase-selective peptide fragment (DEVD), and c). cell-penetrating moiety. The cell-penetrating moiety carries the probe into live cells. Upon entering live cells the caspase-selective peptide fragment is cleaved by a caspase to release the masked fluorophore. The intensity of recovered fluorescence is directly related to the activity of caspase to be measured. Compared to the existing caspase assays in live cells, ApoBrite™ V600 is much more robust, convenient and accurate. ApoBrite™ V600 releases a fluorophore that has a large Stokes shift, and can be well excited with violet laser that is installed most of new flow cytometers. It does not need a DNA interaction to be fluorescent as reported for NucView reagents. It does not inhibit caspase activity as reported for the FMK peptide probes. Although fluorescent FMK peptide inhibitors of caspases are widely used for detecting caspase activities in live cells, this technology has a few severe limitations: a). FMK caspase inhibitors have high cytotoxicity since FMK peptides bind covalently to active caspases; b). The irreversibly covalent binding of FMK peptides to caspases inhibits caspase activities, causing false positive apoptosis; c). FMK assays have extremely high background, and require intensive washings, resulting in very low through put; d). FMK peptides are not stable in aqueous solutions, and have to be used immediately.

Platform


Flow cytometer

Excitation405 nm
Emission570 nm
Instrument specification(s)FL3 channel for 7-AAD staining

Fluorescence microscope

ExcitationDAPI channel
EmissionDAPI channel
Recommended plateBlack wall/clear bottom
Instrument specification(s)TRITC channel for 7-AAD staining

Fluorescence microplate reader

Excitation405 nm
Emission570 nm
Cutoff540 nm
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode

Components


Component A: ApoBrite™ V570 caspase 3/7 substrate1 vial (40 µL, 500 X)
Component B: HBSS 1 bottle (100 mL)
Component C: 500X 7-AAD1 vial (100 µL)

Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cells 
  2. Remove the growth medium, wash cells
  3. Incubate Caspase 3/7 Substrate working solution at  37 ºC for 2 hours 
  4. Wash cells and replace with growth medium
  5. Induce apoptosis
  6. Monitor fluorescence intensity (bottom read mode) at Ex/Em = 405/570 nm (Cutoff = 540 nm), fluorescence microscope with DAPI filter, or flow cytometer with Ex/Em = 405/470 nm filters

Important notes
Thaw all the components at room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

Add 2 μL of 500X ApoBrite™ V570 Caspase 3/7 Substrate stock solution (Component A) into 1 mL HHBS (Component B) to make 1X ApoBrite™ V570 Caspase 3/7 Substrate working solution. 

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Culture cells to a density optimal for apoptosis induction according to your specific induction protocol, but not to exceed 2 x 106 cells/mL for suspension cells. For adherent cells, plate cells at 30,000 - 80,000 cells/well/96-well plate overnight.
         
  2. Remove growth medium, and wash once with HHBS (Component B) or buffer of your choice.

  3. Add 100 µL/well/96 well plate of 1X ApoBrite™ V570 Caspase 3/7 Substrate working solution, and incubate at 37 ºC, 5% CO2 incubator for 2 hours.

  4. Remove 1X ApoBrite™ V570 Caspase 3/7 Substrate working solution and wash once with HHBS (Component B).

  5. Add growth medium back to the ApoBrite™ V570 Caspase 3/7 loaded cells, and treat the cells as desired for apoptosis. Here are a few examples for  inducing apoptosis in cell culture:

    1. Treating Jurkat cells with 2 µg/ml camptothecin for 3 hours.

    2. Treating Jurkat cells with 1 µM staurosporine for 3 hours. Hela cells for 1 hour.

    3. Treating HL-60 cells with 4 µg/ml camptothecin for 4 hours.

    4. Treating HL-60 cells with 1 µM staurosporine for 4 hours.

    5. Treating Hela cells with 1 µM staurosporine for 1 hour. Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.

  6. If desired, label the cells with a DNA stain (such as 7-AAD for dead cells).

  7. Monitor the fluorescence intensity by fluorescence microscopy, flow cytometer, or fluorescence microplate reader at Ex/Em = 405/570 nm (for 7-AAD, Ex/Em = 535/650 nm).

    For flow cytometry (suspension cells): Monitor the fluorescence intensity using Ex/Em = 405/570 nm filters (FL3 channel for 7-AAD staining). Gate on the cells of interest, excluding debris.

    For fluorescence microscope:  Observe cells under a fluorescence microscope using Ex/Em = 405/570 nm or DAPI channel (TRITC channel for 7-AAD staining).

    For fluorescence microplate reader: Monitor the fluorescence intensity (bottom read mode) with a fluorescence plate reader at  Ex/Em = 405/570 nm (Cutoff = 540 nm).

Citations


View all 15 citations: Citation Explorer
Helicobacter pylori secreted protein HP1286 triggers apoptosis in macrophages via TNF-independent and ERK MAPK-dependent pathways
Authors: Tavares, Raquel and Pathak, Sushil Kumar
Journal: Frontiers in Cellular and Infection Microbiology (2017): 58
Promotion of osteointegration under diabetic conditions by tantalum coating-based surface modification on 3-dimensional printed porous titanium implants
Authors: Wang, Lin and Hu, Xiaofan and Ma, Xiangyu and Ma, Zhensheng and Zhang, Yang and Lu, Yizhao and Li, Xiang and Lei, Wei and Feng, Yafei
Journal: Colloids and Surfaces B: Biointerfaces (2016): 440--452
microRNA-186 inhibits cell proliferation and induces apoptosis in human esophageal squamous cell carcinoma by targeting SKP2
Authors: He, Wei and Feng, Jianfang and Zhang, Yan and Wang, Yuanyuan and Zang, Wenqiao and Zhao, Guoqiang
Journal: Laboratory Investigation (2016): 317--324
Hypoxia regulates TRAIL sensitivity of colorectal cancer cells through mitochondrial autophagy.
Authors: Knoll, Gertrud and Bittner, Sebastian and Kurz, Maria and Jantsch, Jonathan and Ehrenschwender, Martin
Journal: Oncotarget (2016)
Role of delta-like ligand-4 in chemoresistance against docetaxel in MCF-7 cells
Authors: Wang, Q and Shi, Y and Butler, HJ and Xue, J and Wang, G and Duan, P and Zheng, H
Journal: Human & Experimental Toxicology (2016): 0960327116650006
Angiopoietins Modulate Survival, Migration, and the Components of the Ang-Tie2 Pathway of Chronic Lymphocytic Leukaemia (CLL) Cells In Vitro
Authors: Palma, Luis Mario Aguirre and Flamme, Hanna and Gerke, Iris and Kreuzer, Karl-Anton
Journal: Cancer Microenvironment (2016): 13--26
t-BHQ Provides Protection against Lead Neurotoxicity via Nrf2/HO-1 Pathway
Authors: Ye, Fang and Li, Xiaoyi and Li, Lili and Yuan, Jing and Chen, Jun
Journal: Oxidative medicine and cellular longevity (2015)
Insulin improves osteogenesis of titanium implants under diabetic conditions by inhibiting reactive oxygen species overproduction via the PI3K-Akt pathway
Authors: Wang, Lin and Zhao, Xiong and Wei, Bo-yuan and Liu, Yi and Ma, Xiang-yu and Wang, Jian and Cao, Peng-chong and Zhang, Yang and Yan, Ya-bo and Lei, Wei and others, undefined
Journal: Biochimie (2015): 85--93
Glucose promotes cell proliferation, glucose uptake and invasion in endometrial cancer cells via AMPK/mTOR/S6 and MAPK signaling
Authors: Han, Jianjun and Zhang, Lu and Guo, Hui and Wysham, Weiya Z and Roque, Dario R and Willson, Adam K and Sheng, Xiugui and Zhou, Chunxiao and Bae-Jump, Victoria L
Journal: Gynecologic oncology (2015): 668--675
IL-7 abrogates the immunosuppressive function of human double-negative T cells by activating Akt/mTOR signaling
Authors: Allgäuer, Andrea and Schreiner, Elisabeth and Ferrazzi, Fulvia and Ekici, Arif B and Gerbitz, Armin and Mackensen, Andreas and Völkl, Simon
Journal: The Journal of Immunology (2015): 3139--3148