Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*
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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
Correction Factor (260 nm) | 0.21 |
Correction Factor (280 nm) | 0.11 |
Extinction coefficient (cm -1 M -1) | 750001 |
Excitation (nm) | 491 |
Emission (nm) | 516 |
Quantum yield | 0.91 |
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: Annexin-V Staining, Apoptosis and Necrosis, Cell Viability Assays, Cellular Processes, Flow Cytometry Reagents
Correction Factor (260 nm) 0.21 | Correction Factor (280 nm) 0.11 | Extinction coefficient (cm -1 M -1) 750001 | Excitation (nm) 491 | Emission (nm) 516 | Quantum yield 0.91 |
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 the translocation of phosphatidylserine (PS). In apoptosis, PS is transferred to the outer leaflet of the plasma membrane. The appearance of phosphatidylserine on the cell surface is a universal indicator of the initial/intermediate stages of cell apoptosis and can be detected before morphological changes can be observed. This kit uses a fluorescent Annexin V that specifically binds PS. Annexin V conjugates have been demonstrated to selectively bind PS. This particular assay kit is optimized to monitor cell apoptosis using a flow cytometer with the FITC channel (green fluorescence).
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds (200 µL/sample)
- Add Annexin V-iFluor™ 488 stock solution
- Incubate at room temperature for 30 - 60 mintues
- Analyze cells using flow cytometer with 530/30 nm filter (FITC channel)
Important notes
Thaw 100X Propidium Iodide (Component C) at room temperature before starting the experiment.
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells with test compounds for a desired period of time (4 - 6 hours for Jurkat cells treated with camptothecin) to induce apoptosis. Note: Annexin V flow cytometric analysis on adherent cells is not routinely tested since specific membrane damage may occur during cell detachment or harvesting. However, methods for utilizing Annexin V for flow cytometry on adherent cell types have been previously reported by Casiola-Rosen et al. and van Engelend et al.
- Centrifuge the cells to get 1 - 5 × 105 cells/tube.
- Resuspend cells in 200 µL of Assay Buffer (Component B).
- Add 2 µL of Annexin V-iFluor™ 488 (Component A) into the cells.
- Optional: Add 2 µL of 100X Propidium Iodide (Component C) for necrosis cells.
- Incubate at room temperature for 30 to 60 minutes, protected from light.
- Optional: Add 200 to 300 µL of Assay Buffer (Component B) to increase volume before analyzing the cells with a flow cytometer.
- Monitor the fluorescence intensity of Annexin V-iFluor™ 488 using a flow cytometer with 530/30 nm filter (FITC channel). Measure the cell viability using the 610/20 nm filter (PE-Texas Red channel) when propidium iodide is added into the cells.
Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Correction Factor (260 nm) | 0.21 |
Correction Factor (280 nm) | 0.11 |
Extinction coefficient (cm -1 M -1) | 750001 |
Excitation (nm) | 491 |
Emission (nm) | 516 |
Quantum yield | 0.91 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Orange Fluorescence Optimized for Flow Cytometry* | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Red Fluorescence Optimized for Flow Cytometry* | 588 | 604 | 1800001 | 0.531 | 0.05 | 0.04 |
Cell Meter™ Annexin V Binding Apoptosis Assay Kit *Deep Red Fluorescence Optimized for Flow Cytometry* | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
Images

Figure 1. The detection of binding activity of Annexin V-iFluor® 488 and phosphatidylserine in Jurkat cells. Jurkat cells were treated without (Blue) or with 20 µM camptothecin (Red) in a 37 °C, 5% CO2 incubator for 4-5 hours, and then dye loaded with Annexin V-iFluor® 488 for 30 minutes. The fluorescence intensity of Annexin V-iFluor® 488 was measured with a FACSCalibur (Becton Dickinson) flow cytometer using the FL1 channel.

Figure 2. Effect of FSS on apoptosis and necrosis in tubular cells. Confluent monolayers of HK-2 cells were submitted to FSS 0 (static) or FSS 0.5 Pa (FSS 0.5) for 48h. A/ Cells were stained with Annexin-V and then immediately subjected to analysis of phosphatidylserine externalization (Annexin-V fluorescence, X-axis) and Propidium Iodure (PI) uptake (PI fluorescence, Y-axis) using flow cytometry. Living, early apoptotic or necrotic (primary or secondary) cells were distinguished by the criteria of Annexin-V−/PI−(bottom left quadrant), Annexin-V+/PI− (bottom right quadrant) and Annexin-V+/PI+ (upper right quadrant), respectively. B/ Proportions of early apoptosis and necrosis cells were quantified and results are expressed as a percentage of the total population of cells. Data represent mean ± SEM of 7 experiments. *HK-2 cells were assessed for apoptosis and necrosis using Cell Meter Annexin V Binding Apoptosis Assay Kit (AAT Bioquest), according to the manufacturer's instructions. Source: Graph from Shear Stress-Induced Alteration of Epithelial Organization in Human Renal Tubular Cells by Damien Maggiorani, et al., PLoS ONE, July 2015.
Citations
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CCT6A knockdown suppresses osteosarcoma cell growth and Akt pathway activation in vitro
Authors: Zeng, Weiquan and Wu, Meizhu and Cheng, Ying and Liu, Liya and Han, Yuying and Xie, Qiurong and Li, Jiapeng and Wei, Lihui and Fang, Yi and Chen, Youqin and others,
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Authors: Lala, Trisha and Doan, Juleva K and Takatsu, Hiroyuki and Hartzell, H Criss and Shin, Hye-Won and Hall, Randy A
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Atorvastatin attenuates pulmonary fibrosis in mice and human lung fibroblasts, by the regulation of myofibroblast differentiation and apoptosis
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Authors: Yildirim, Merve and Kayalar, Ozgecan and Atahan, Ersan and Oztay, Fusun
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Itraconazole improves survival outcomes in patients with colon cancer by inducing autophagic cell death and inhibiting transketolase expression
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Journal: Eneuro (2020)
Combined Treatments of Magnetic Intra-Lysosomal Hyperthermia with Doxorubicin Promotes Synergistic Anti-Tumoral Activity.
Authors: El, D Hajj Diab and Clerc, P and Serhan, N and Fourmy, D and Gigoux, V
Journal: Nanomaterials (Basel, Switzerland) (2018)
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Journal: Nanomaterials (Basel, Switzerland) (2018)
In situ polymerizable hydrogel incorporated with specific pathogen-free porcine platelet-rich plasma for the reconstruction of the corneal endothelium
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Journal: Journal of the Taiwan Institute of Chemical Engineers (2017)
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Journal: Journal of the Taiwan Institute of Chemical Engineers (2017)
Novel regulations of MEF2-A, MEF2-D, and CACNA1S in the functional incompetence of adipose-derived mesenchymal stem cells by induced indoxyl sulfate in chronic kidney disease
Authors: Do, Duyen Thi and Phan, Nam Nhut and Wang, Chih-Yang and Sun, Zhengda and Lin, Yen-Chang
Journal: Cytotechnology (2016): 2589--2604
Authors: Do, Duyen Thi and Phan, Nam Nhut and Wang, Chih-Yang and Sun, Zhengda and Lin, Yen-Chang
Journal: Cytotechnology (2016): 2589--2604
Shear stress-induced alteration of epithelial organization in human renal tubular cells
Authors: Maggiorani, Damien and Dissard, Romain and Belloy, Marcy and Saulnier-Blache, Jean-Sébastien and Casemayou, Audrey and Ducasse, Laure and Grès, S and ra , undefined and Bellière, Julie and Caubet, Cécile and Basc, undefined and s, Jean-Loup and others, undefined
Journal: PloS one (2015): e0131416
Authors: Maggiorani, Damien and Dissard, Romain and Belloy, Marcy and Saulnier-Blache, Jean-Sébastien and Casemayou, Audrey and Ducasse, Laure and Grès, S and ra , undefined and Bellière, Julie and Caubet, Cécile and Basc, undefined and s, Jean-Loup and others, undefined
Journal: PloS one (2015): e0131416
Shear stress-induced alteration of epithelial organization in human renal tubular cells
Authors: Maggiorani, Damien and Dissard, Romain and Belloy, Marcy and Saulnier-Blache, Jean-S{\'e}bastien and Casemayou, Audrey and Ducasse, Laure and Gr{\`e}s, Sandra and Belli{\`e}re, Julie and Caubet, C{\'e}cile and Bascands, Jean-Loup and others,
Journal: PLoS One (2015): e0131416
Authors: Maggiorani, Damien and Dissard, Romain and Belloy, Marcy and Saulnier-Blache, Jean-S{\'e}bastien and Casemayou, Audrey and Ducasse, Laure and Gr{\`e}s, Sandra and Belli{\`e}re, Julie and Caubet, C{\'e}cile and Bascands, Jean-Loup and others,
Journal: PLoS One (2015): e0131416
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