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Description
The Annexin V-YSFluor™ 647/PI Apoptosis Detection Kit uses Annexin V conjugated to YSFluor™ 647 as a probe to detect early-stage apoptosis in cells. The assay can be analyzed by flow cytometry or other fluorescence detection instruments.
Principle of Detection:In healthy viable cells, phosphatidylserine (PS) is localized on the inner leaflet of the plasma membrane. However, during early apoptosis, PS is translocated to the outer surface of the cell membrane and becomes exposed to the extracellular environment. Annexin V is a 35–36 kDa Ca²⁺-dependent phospholipid-binding protein with high affinity for PS. It binds specifically to PS exposed on the outer membrane of early apoptotic cells.
Additionally, this kit includes Propidium Iodide (PI), a nucleic acid dye used to distinguish viable early apoptotic cells from late apoptotic or necrotic cells. PI cannot penetrate intact membranes of live or early apoptotic cells but readily enters cells with compromised membranes—such as late apoptotic or necrotic cells—and stains their nuclei red.
Therefore, when Annexin V and PI are used together:
Viable cells are Annexin V⁻/PI⁻
Early apoptotic cells are Annexin V⁺/PI⁻
Late apoptotic and necrotic cells are Annexin V⁺/PI⁺ (double-positive)
This kit requires analysis by flow cytometry.
Features
- Rapid and Convenient Detection – Simple workflow enables fast assessment of apoptosis in cultured cells without complex procedures.
- High Accuracy – Dual staining with Annexin V-YSFluor™ 647 and Propidium Iodide (PI) allows precise discrimination between early apoptotic, late apoptotic, necrotic, and live cells.
- Sensitive and Reliable – Optimized fluorophore conjugation and buffer formulation ensure strong signal with minimal background, suitable for flow cytometry and fluorescence microscopy.
- Versatile – Compatible with a wide range of cell types, including adherent and suspension cells.
Components
|
Components No. |
Name |
40304ES20 (20 T) |
40304ES50 (50 T) |
40304ES60 (100 T) |
|
40304-A |
Annexin V-YSFluor™ 647 |
100 μL |
250 μL |
500 μL |
|
40304-B |
PI Staining Solution (20 μg/mL) |
200 μL |
500 μL |
1.0 mL |
|
40304-C |
1× Binding Buffer |
10 mL |
25 mL |
50 mL |
Storage
This product should be stored at -25℃~-15℃ for one year.
Documents:
Safety Data Sheet
Manuals
40304_Manual_Ver.EN20251111.pdf
Publications Using This Product
[1] Qu S, Jiao Z, Lu G, et al. PD-L1 lncRNA splice isoform promotes lung adenocarcinoma progression via enhancing c-Myc activity. Genome Biol. 2021;22(1):104. Published 2021 Apr 13. doi:10.1186/s13059-021-02331-0(IF:13.583)
[2] Wang J, Du X, Wang X, et al. Tumor-derived miR-378a-3p-containing extracellular vesicles promote osteolysis by activating the Dyrk1a/Nfatc1/Angptl2 axis for bone metastasis. Cancer Lett. 2022;526:76-90. doi:10.1016/j.canlet.2021.11.017(IF:8.679)
[3] Xie J, Xu W, Wu Y, Niu B, Zhang X. Macroporous organosilicon nanocomposites co-deliver Bcl2-converting peptide and chemotherapeutic agent for synergistic treatment against multidrug resistant cancer. Cancer Lett. 2020;469:340-354. doi:10.1016/j.canlet.2019.10.018(IF:6.508)
[4] Li S, Li X, Chen F, et al. Nobiletin mitigates hepatocytes death, liver inflammation, and fibrosis in a murine model of NASH through modulating hepatic oxidative stress and mitochondrial dysfunction. J Nutr Biochem. 2022;100:108888. doi:10.1016/j.jnutbio.2021.108888(IF:6.048)
[5] Han L, Wu Y, Liu F, Zhang H. eIF4A1 Inhibitor Suppresses Hyperactive mTOR-Associated Tumors by Inducing Necroptosis and G2/M Arrest. Int J Mol Sci. 2022;23(13):6932. Published 2022 Jun 22. doi:10.3390/ijms23136932(IF:5.924)
[6] Gao F, Wang Q, Zhang C, et al. RNA methyltransferase METTL3 induces intrinsic resistance to gefitinib by combining with MET to regulate PI3K/AKT pathway in lung adenocarcinoma. J Cell Mol Med. 2021;25(5):2418-2425. doi:10.1111/jcmm.16114(IF:5.310)
[7] Ren L, Hu L, Zhang Y, et al. Cataract-Causing S93R Mutant Destabilized Structural Conformation of βB1 Crystallin Linking With Aggregates Formation and Cellular Viability. Front Mol Biosci. 2022;9:844719. Published 2022 Mar 14. doi:10.3389/fmolb.2022.844719(IF:5.246)
[8] Ding M, Weng C, Fan S, Cao Q, Lu Z. Purkinje Cell Degeneration and Motor Coordination Deficits in a New Mouse Model of Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay. Front Mol Neurosci. 2017;10:121. Published 2017 May 1. doi:10.3389/fnmol.2017.00121(IF:5.076)
[9] Cai C, Dang W, Liu S, et al. Anthrax toxin receptor 1/tumor endothelial marker 8 promotes gastric cancer progression through activation of the PI3K/AKT/mTOR signaling pathway. Cancer Sci. 2020;111(4):1132-1145. doi:10.1111/cas.14326(IF:4.966)
[10] Li X, Yao Q, Huang J, et al. Morin Hydrate Inhibits TREM-1/TLR4-Mediated Inflammatory Response in Macrophages and Protects Against Carbon Tetrachloride-Induced Acute Liver Injury in Mice. Front Pharmacol. 2019;10:1089. Published 2019 Sep 20. doi:10.3389/fphar.2019.01089(IF:3.845)
[11] Jiang Y, Du M, Wu M, et al. Phosphatidic Acid Improves Reprogramming to Pluripotency by Reducing Apoptosis. Stem Cells Dev. 2016;25(1):43-54. doi:10.1089/scd.2015.0159(IF:3.727)
[12] Zhang Q, Wu C, Fan Y, et al. Nucleic acid-targeted pathogen reduction technique in red blood cells by UV-generated oxygen radicals for optimising recipient safety. Transfus Med. 2020;30(1):51-60. doi:10.1111/tme.12654(IF:1.900)
During early apoptosis, phosphatidylserine (PS) translocates from the inner to the outer leaflet of the plasma membrane before membrane integrity is lost. Annexin V binds PS with high affinity in a calcium-dependent manner, enabling early-apoptosis detection. Co-staining with propidium iodide (PI) discriminates: Annexin V⁺/PI⁻ = early apoptosis; Annexin V⁺/PI⁺ = late apoptosis/necrosis; Annexin V⁻/PI⁻ = viable cells.
TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) detects DNA strand breaks — a late-stage hallmark of apoptosis — by enzymatically labeling 3′-OH termini with fluorescent dUTP. TUNEL is preferred for detecting apoptosis in fixed cells, tissue sections, or when intracellular markers are needed (e.g., co-staining with nuclear markers). Annexin V is suitable for live, intact cells analyzed by flow cytometry or fluorescence microscopy.
Channel requirements depend on the kit format: FITC variants require a 488 nm laser with 530/30 nm BP filter; YSFluor 488 variants are similar; YSFluor 647 requires a 633/647 nm (red) laser with 660/20 nm or 670 LP filter. Confirm your cytometer's laser and filter configuration matches the kit's excitation/emission profile. PI (propidium iodide) is typically detected in the PE or PerCP channel (488 nm excitation, 610–620 nm emission).
Avoid mechanical or chemical stress during harvest — use gentle enzymatic dissociation (Accutase preferred over trypsin) and keep cells at 4°C during staining. Wash with cold Annexin V Binding Buffer (calcium-containing) immediately before staining. Do not fix cells before Annexin V staining — fixation disrupts membrane phospholipid asymmetry. Process and analyze samples within 1 hour of staining to prevent signal drift.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
