Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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Spectral properties
Extinction coefficient (cm -1 M -1) | 700000 |
Excitation (nm) | 651 |
Emission (nm) | 660 |
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 |
Related products
Overview | ![]() ![]() |
See also: Annexin-V Staining, Flow Cytometry Reagents
Extinction coefficient (cm -1 M -1) 700000 | Excitation (nm) 651 | Emission (nm) 660 |
Annexin V may be conjugated to fluorochromes including APC. This format retains its high affinity for phosphatidylserine (PS) and thus serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. Since externalization of PS occurs in the earlier stages of apoptosis, APC Annexin V staining can identify apoptosis at an earlier stage than assays based on nuclear changes such as DNA fragmentation. APC Annexin V staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from either apoptotic or necrotic processes. Therefore, staining with APC Annexin V is typically used in conjunction with a vital dye such as propidium iodide (PI) or 7-Amino-Actinomycin (7-AAD) to allow the investigator to identify early apoptotic cells.
Platform
Flow cytometer
Excitation | 640 nm laser |
Emission | 660/20 nm filter |
Instrument specification(s) | APC channel |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds (200 µL/sample)
- Add APC-Annexin V assay solution
- Incubate at room temperature for 20 - 60 minutes
- Analyze cells using flow cytometer with 660/20 nm filter (APC channel)
Important notes
Thaw 100X Propidium Iodide (Component C) at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1. APC-Annexin V stock solution (100X):
Add 200 µL PBS with 0.2% BSA into the vial of APC-Annexin V conjugate (Component A) and mix well to make 100X APC-Annexin V stock solution. Note: Store the reconstituted 100X APC-Annexin V stock solution at 4 oC. Do Not Freeze.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells with test compounds for a desired period of time (4-6 hours for Jurkat cells treated with staurosporine) 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 100X APC-Annexin V stock solution into the cells.
- Optional: Add 2 µL of 100X Propidium Iodide (Component C) into the cells for necrosis cells.
- Incubate at room temperature for 20 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 APC-Annexin V using a flow cytometer with 660/20 nm filter (APC channel). Measure the cell viability using 610/20 nm filter (PE-Texas Red channel) when propidium iodide is added into the cells.
Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Extinction coefficient (cm -1 M -1) | 700000 |
Excitation (nm) | 651 |
Emission (nm) | 660 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (280 nm) |
Cell Meter™ PE-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry* | 565 | 574 | 1960000 | 0.82 | 0.16 |
Cell Meter™ FITC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry* | 491 | 516 | 73000 | 0.92 | 0.35 |
Images

Figure 1. The detection of binding activity of APC-Annexin V to phosphatidylserine in Jurkat cells with Cell Meter™ APC-Annexin V Binding Apoptosis Assay Kit. Jurkat cells were treated without (Blue) or with 1 µM staurosporine (Red) in a 37 °C, 5% CO2 incubator for ~4 hours, and then dye loaded with APC-Annexin V for 30 minutes. The fluorescence intensity of APC-Annexin V was measured with a FACSCalibur (Becton Dickinson) flow cytometer using the FL4 channel.
Citations
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Journal: Frontiers in Bioscience-Landmark (2023): 89
Authors: Gong, Huiyuan and Ma, Chao and Li, Xiaojun and Zhang, Xueying and Zhang, Linxiang and Chen, Pengfei and Wang, Wei and Hu, Yannan and Huang, Ting and Wu, Nan and others,
Journal: Frontiers in Bioscience-Landmark (2023): 89
BRAF D594A mutation defines a unique biological and immuno-modulatory subgroup associated with functional CD8+ T cell infiltration in colorectal cancer
Authors: Li, Wenjing and Zhao, Chenyi and Li, Wenhui and Gong, Yang and Ma, Kaili and Lu, Yujie and Liu, Xiaowei and Zhang, Lianjun and Guo, Feng
Journal: (2023)
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Th22/IL-22 mediates the progression of HBV-related hepatocellular carcinoma via STAT3
Authors: Zhang, Jia and Liu, Zhou and Liu, Lingpeng and Huang, Mingwen and Huang, Yong
Journal: Cytotechnology (2022): 203--216
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Effects of Y-27632 on the osteogenic and adipogenic potential of human dental pulp stem cells in vitro
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Journal: Human \& Experimental Toxicology (2022): 09603271221089003
Knockdown of CENPK inhibits cell growth and facilitates apoptosis via PTEN-PI3K-AKT signalling pathway in gastric cancer
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Journal: Journal of Cellular and Molecular Medicine (2021): 8890--8903
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Journal: Journal of Cellular and Molecular Medicine (2021): 8890--8903
FIGNL1 promotes non-small cell lung cancer cell proliferation
Authors: Li, Miao and Rui, Yan and Peng, Wenjia and Hu, Junfeng and Jiang, Anbang and Yang, Zeyu and Huang, Linian
Journal: International journal of oncology (2020): 83--99
Authors: Li, Miao and Rui, Yan and Peng, Wenjia and Hu, Junfeng and Jiang, Anbang and Yang, Zeyu and Huang, Linian
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Babao Dan induces gastric cancer cell apoptosis via regulating MAPK and NF-$\kappa$B signaling pathways
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Journal: Journal of International Medical Research (2019): 5106--5119
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CHIP functions as an oncogene by promoting colorectal cancer metastasis via activation of MAPK and AKT signaling and suppression of E-cadherin
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Ubiquitin ligase CHIP functions as an oncogene and activates the AKT signaling pathway in prostate cancer
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Authors: Sarvi, Sana and Crispin, Richard and Lu, Yuting and Zeng, Lifan and Hurley, Thomas D and Houston, Douglas R and von Kriegsheim, Alex and Chen, Che-Hong and Mochly-Rosen, Daria and Ranzani, Marco and others, undefined
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Authors: Sarvi, Sana and Crispin, Richard and Lu, Yuting and Zeng, Lifan and Hurley, Thomas D and Houston, Douglas R and von Kriegsheim, Alex and Chen, Che-Hong and Mochly-Rosen, Daria and Ranzani, Marco and others, undefined
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