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Description
The Annexin V-YSFluor™ 488/PI Apoptosis Detection Kit utilizes Annexin V conjugated to YSFluor™ 488 as a fluorescent probe to detect early-stage apoptosis. The assay can be analyzed by flow cytometry or other fluorescence detection platforms.
Principle of Detection:In healthy, viable cells, phosphatidylserine (PS) is localized to the inner leaflet of the plasma membrane. During early apoptosis, PS is translocated to the outer surface of the cell membrane, becoming exposed to the extracellular environment. Annexin V is a 35–36 kDa Ca²⁺-dependent phospholipid-binding protein with high affinity for PS. Fluorescently labeled Annexin V-YSFluor™ 488 binds specifically to PS exposed on the surface of early apoptotic cells.
This kit also includes Propidium Iodide (PI), a nucleic acid dye used to distinguish early apoptotic cells from late apoptotic or necrotic cells. PI is membrane-impermeant and cannot enter live or early apoptotic cells with intact membranes. However, it readily penetrates cells with compromised membranes—such as late apoptotic or necrotic cells—and stains their nuclei red.
When used together: Viable cells: Annexin V⁻ / PI⁻; Early apoptotic cells: Annexin V⁺ / PI⁻; Late apoptotic or necrotic cells: Annexin V⁺ / PI⁺ (dual-positive).
Features
- Rapid and reliable detection: Enables fast and accurate discrimination between live, early apoptotic, late apoptotic, and necrotic cells.
- Dual-color staining: Combines Annexin V-YSFluor™ 488 for early apoptosis (green) and propidium iodide (PI) for late apoptosis or necrosis (red).
- High sensitivity and low background: Provides clear, distinct population separation in flow cytometry or fluorescence microscopy.
- Wide compatibility: Applicable to both adherent and suspension cells from various mammalian species.
- Simple protocol: One-step staining procedure without cell fixation, preserving cell morphology and viability.
Components
|
Components No. |
Name |
40305ES20 (20 T) |
40305ES50 (50 T) |
40305ES60 (100 T) |
|
40305-A |
Annexin V-YSFluor™ 488 |
100 μL |
250 μL |
500 μL |
|
40305-B |
PI Staining Solution (20 μg/mL) |
200 μL |
500 μL |
1.0 mL |
|
40305-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
40305_Manual_Ver.EN20251111.pdf
Publications Using This Product
[1] Zhang M, Weng Y, Cao Z, et al. ROS-Activatable siRNA-Engineered Polyplex for NIR-Triggered Synergistic Cancer Treatment. ACS Appl Mater Interfaces. 2020;12(29):32289-32300. doi:10.1021/acsami.0c06614(IF:8.758)
[2] Duan Z , Luo Q , Gu L , et al. A co-delivery nanoplatform for a lignan-derived compound and perfluorocarbon tuning IL-25 secretion and the oxygen level in tumor microenvironments for meliorative tumor radiotherapy. Nanoscale. 2021;13(32):13681-13692. doi:10.1039/d1nr03738b(IF:7.790)
[3] Tang L, Zhang C, Lu L, et al. Melatonin Maintains Inner Blood-Retinal Barrier by Regulating Microglia via Inhibition of PI3K/Akt/Stat3/NF-κB Signaling Pathways in Experimental Diabetic Retinopathy. Front Immunol. 2022;13:831660. Published 2022 Mar 15. doi:10.3389/fimmu.2022.831660(IF:7.561)
[4] Xie H, Zhang C, Liu D, et al. Erythropoietin protects the inner blood-retinal barrier by inhibiting microglia phagocytosis via Src/Akt/cofilin signalling in experimental diabetic retinopathy. Diabetologia. 2021;64(1):211-225. doi:10.1007/s00125-020-05299-x(IF:7.518)
[5] Yue C, Yang Y, Song J, et al. Mitochondria-targeting near-infrared light-triggered thermosensitive liposomes for localized photothermal and photodynamic ablation of tumors combined with chemotherapy. Nanoscale. 2017;9(31):11103-11118. doi:10.1039/c7nr02193c(IF:7.367)
[6] Yu H, Yang X, Xiao X, et al. Human Adipose Mesenchymal Stem Cell-derived Exosomes Protect Mice from DSS-Induced Inflammatory Bowel Disease by Promoting Intestinal-stem-cell and Epithelial Regeneration. Aging Dis. 2021;12(6):1423-1437. Published 2021 Sep 1. doi:10.14336/AD.2021.0601(IF:6.745)
[7] Huang J, Liu Y, Chen JX, et al. Harmine is an effective therapeutic small molecule for the treatment of cardiac hypertrophy. Acta Pharmacol Sin. 2022;43(1):50-63. doi:10.1038/s41401-021-00639-y(IF:6.150)
[8] Wang X, Zhang R, Wu T, et al. Successive treatment with naltrexone induces epithelial-mesenchymal transition and facilitates the malignant biological behaviors of bladder cancer cells. Acta Biochim Biophys Sin (Shanghai). 2021;53(2):238-248. doi:10.1093/abbs/gmaa169(IF:3.848)
[9] Qu B, Han X, Zhao L, Zhang F, Gao Q. Relationship of HIF‑1α expression with apoptosis and cell cycle in bone marrow mesenchymal stem cells from patients with myelodysplastic syndrome. Mol Med Rep. 2022;26(1):239. doi:10.3892/mmr.2022.12755(IF:2.952)
[10] Fan W, Gao X, Ding C, et al. Estrogen receptors participate in carcinogenesis signaling pathways by directly regulating NOD-like receptors. Biochem Biophys Res Commun. 2019;511(2):468-475. doi:10.1016/j.bbrc.2019.02.085(IF:2.705)
[11] Feng Z, Zhu Z, Chen W, Bai Y, Hu D, Cheng J. Chloride intracellular channel 4 participate in the protective effect of Ginkgolide B in MPP+ injured MN9D cells: insight from proteomic analysis. Clin Proteomics. 2020;17:32. Published 2020 Sep 5. doi:10.1186/s12014-020-09295-6(IF:2.568)
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.
