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Description
During apoptosis, cells activate certain DNA endonucleases that cleave genomic DNA between nucleosomes. When DNA is extracted from apoptotic cells and analyzed by electrophoresis, a characteristic DNA ladder of ~180–200 bp fragments can be observed.
The TUNEL (TdT-mediated dUTP Nick-End Labeling) Apoptosis Detection Kit (YSFluor™ 488) is designed to detect DNA fragmentation in the nuclei of cells during late-stage apoptosis. The principle is based on the enzymatic activity of Terminal Deoxynucleotidyl Transferase (TdT), which catalyzes the incorporation of YSFluor™ 488-12-dUTP into the 3′-hydroxyl (3′-OH) ends of fragmented DNA. The labeled DNA can then be visualized using a fluorescence microscope or analyzed by flow cytometry.
YSFluor™ 488 offers higher photostability and stronger fluorescence intensity compared to conventional dyes, resulting in brighter labeling and enhanced resistance to photobleaching.
This kit is broadly applicable and can be used to detect apoptosis in:
Formalin-fixed paraffin-embedded (FFPE) tissue sections; Frozen tissue sections; Cultured adherent or suspension cells.
Features
- Broad applicability: Suitable for apoptosis detection in frozen or paraffin-embedded tissue sections, as well as in adherent and suspension cell cultures.
- High sensitivity and specificity: Accurately distinguishes apoptotic cells from necrotic or normal cells through fluorescent labeling of DNA breaks.
- Bright and stable fluorescence: YSFluor™ 488 provides strong green fluorescence with excellent photostability, ensuring clear and reproducible imaging.
- Simple and rapid protocol: Streamlined workflow for convenient use in various experimental setups.
- Compatible with multiple detection platforms: Applicable to fluorescence microscopy, flow cytometry, and high-content imaging systems.
Components
|
Components No. |
Name |
40307ES20 (20 T) |
40307ES50 (50 T) |
40307ES60 (100 T) |
|
40307-A |
5× Equilibration Buffer |
750 μL |
1.25 mL × 2 |
1.25 mL × 3 |
|
40307-B |
YSFluor™ 488-12-dUTP Labeling Mix |
100 μL |
250 μL |
250 μL × 2 |
|
40307-C |
Recombinant TdT Enzyme |
20 μL |
50 μL |
50 μL × 2 |
|
40307-D |
Proteinase K (2 mg/mL) |
40 μL |
100 μL |
100 μL × 2 |
|
40307-E |
DNase I (1 U/μL) |
5 μL |
12.5 μL |
25 μL |
|
40307-F |
10× DNase I Buffer with MgCl₂ |
100 μL |
250 μL |
500 μL |
Storage
Shipped on wet ice.
Store the entire kit at –25~-15°C. The YSFluor™ 488-12-dUTP Labeling Mix must be protected from light and stored at –20°C. The kit is stable for 1 year under these conditions.
Notes
1. Users must prepare the following reagents:
PBS for cell washing
Antifade mounting medium for microscopy
4% paraformaldehyde (in PBS) for fixation
2. For safety and hygiene, wear a lab coat and disposable gloves during handling.
3. This product is for research use only!
Documents:
Publications Using This Product
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[2] Li Z, Zhang X, Xie S, et al. H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis [published online ahead of print, 2022 Jun 23]. Nucleic Acids Res. 2022;50(12):6786-6800. doi:10.1093/nar/gkac533(IF:16.971)
[3] Jin X, Wang K, Wang L, et al. RAB7 activity is required for the regulation of mitophagy in oocyte meiosis and oocyte quality control during ovarian aging. Autophagy. 2022;18(3):643-660. doi:10.1080/15548627.2021.1946739(IF:16.016)
[4] Chen Y, Wang Y, Chen J, et al. The SUN1-SPDYA interaction plays an essential role in meiosis prophase I. Nat Commun. 2021;12(1):3176. Published 2021 May 26. doi:10.1038/s41467-021-23550-w(IF:14.919)
[5] Liang W, Peng X, Li Q, et al. FAM3D is essential for colon homeostasis and host defense against inflammation associated carcinogenesis. Nat Commun. 2020;11(1):5912. Published 2020 Nov 20. doi:10.1038/s41467-020-19691-z(IF:12.121)
[6] Zhang YJ, Zhang M, Zhao X, et al. NAD+ administration decreases microvascular damage following cardiac ischemia/reperfusion by restoring autophagic flux. Basic Res Cardiol. 2020;115(5):57. Published 2020 Aug 10. doi:10.1007/s00395-020-0817-z(IF:11.981)
[7] Wang Y, Chen Y, Chen J, et al. The meiotic TERB1-TERB2-MAJIN complex tethers telomeres to the nuclear envelope. Nat Commun. 2019;10(1):564. Published 2019 Feb 4. doi:10.1038/s41467-019-08437-1(IF:11.878)
[8] Yang B, Li L, Tong G, et al. Circular RNA circ_001422 promotes the progression and metastasis of osteosarcoma via the miR-195-5p/FGF2/PI3K/Akt axis. J Exp Clin Cancer Res. 2021;40(1):235. Published 2021 Jul 16. doi:10.1186/s13046-021-02027-0(IF:11.161)
[9] 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)
[10] Zhou K, Zheng Z, Li Y, et al. TFE3, a potential therapeutic target for Spinal Cord Injury via augmenting autophagy flux and alleviating ER stress. Theranostics. 2020;10(20):9280-9302. Published 2020 Jul 23. doi:10.7150/thno.46566(IF:8.579)
[11] Yu H, Yang W, Huang J, et al. GPR120 induces regulatory dendritic cells by inhibiting HK2-dependent glycolysis to alleviate fulminant hepatic failure. Cell Death Dis. 2021;13(1):1. Published 2021 Dec 16. doi:10.1038/s41419-021-04394-0(IF:8.469)
[12] Zhang H, Hao LZ, Pan JA, et al. Microfluidic fabrication of inhalable large porous microspheres loaded with H2S-releasing aspirin derivative for pulmonary arterial hypertension therapy. J Control Release. 2021;329:286-298. doi:10.1016/j.jconrel.2020.11.060(IF:7.727)
[13] Mao G, Xin D, Wang Q, Lai D. Sodium molybdate inhibits the growth of ovarian cancer cells via inducing both ferroptosis and apoptosis. Free Radic Biol Med. 2022;182:79-92. doi:10.1016/j.freeradbiomed.2022.02.023(IF:7.376)
[14] Xu T, Ding W, Ao X, et al. ARC regulates programmed necrosis and myocardial ischemia/reperfusion injury through the inhibition of mPTP opening. Redox Biol. 2019;20:414-426. doi:10.1016/j.redox.2018.10.023(IF:7.126)
[15] Mao Y, Du J, Chen X, et al. Maltol Promotes Mitophagy and Inhibits Oxidative Stress via the Nrf2/PINK1/Parkin Pathway after Spinal Cord Injury. Oxid Med Cell Longev. 2022;2022:1337630. Published 2022 Feb 1. doi:10.1155/2022/1337630(IF:6.543)
[16] Song S, Hou X, Zhang W, et al. Specific bFGF targeting of KIM-1 in ischemic kidneys protects against renal ischemia-reperfusion injury in rats. Regen Biomater. 2022;9:rbac029. Published 2022 May 12. doi:10.1093/rb/rbac029(IF:6.353)
[17] Li Y, Zhang J, Zhou K, et al. Elevating sestrin2 attenuates endoplasmic reticulum stress and improves functional recovery through autophagy activation after spinal cord injury. Cell Biol Toxicol. 2021;37(3):401-419. doi:10.1007/s10565-020-09550-4(IF:6.284)
[18] Wu X, Zhang T, Lyu P, et al. Traditional Chinese Medication Qiliqiangxin Attenuates Diabetic Cardiomyopathy via Activating PPARγ. Front Cardiovasc Med. 2021;8:698056. Published 2021 Jul 16. doi:10.3389/fcvm.2021.698056(IF:6.050)
[19] Pan JA, Tang Y, Yu JY, et al. miR-146a attenuates apoptosis and modulates autophagy by targeting TAF9b/P53 pathway in doxorubicin-induced cardiotoxicity. Cell Death Dis. 2019;10(9):668. Published 2019 Sep 11. doi:10.1038/s41419-019-1901-x(IF:5.959)
[20] Zhang Y, He Z, Li Y, et al. Tumor cell membrane-derived nano-Trojan horses encapsulating phototherapy and chemotherapy are accepted by homologous tumor cells. Mater Sci Eng C Mater Biol Appl. 2021;120:111670. doi:10.1016/j.msec.2020.111670(IF:5.880)
[21] Kang H, Dong Y, Peng R, et al. Inhibition of IRE1 suppresses the catabolic effect of IL-1β on nucleus pulposus cell and prevents intervertebral disc degeneration in vivo. Biochem Pharmacol. 2022;197:114932. doi:10.1016/j.bcp.2022.114932(IF:5.858)
[22] Jin J, Ma Y, Tong X, et al. Metformin inhibits testosterone-induced endoplasmic reticulum stress in ovarian granulosa cells via inactivation of p38 MAPK [published correction appears in Hum Reprod. 2020 Aug 1;35(8):1947-1948]. Hum Reprod. 2020;35(5):1145-1158. doi:10.1093/humrep/deaa077(IF:5.733)
[23] Huang CX, Jiang ZX, Du DY, Zhang ZM, Liu Y, Li YT. The MFF-SIRT1/3 axis, regulated by miR-340-5p, restores mitochondrial homeostasis of hypoxia-induced pulmonary artery smooth muscle cells. Lab Invest. 2022;102(5):515-523. doi:10.1038/s41374-022-00730-w(IF:5.662)
[24] Zhang S, Xing M, Chen G, Tong L, Zhang H, Du D. Up-regulation of miR-335 and miR-674-3p in the rostral ventrolateral medulla contributes to stress-induced hypertension. J Neurochem. 2022;161(5):387-404. doi:10.1111/jnc.15589(IF:5.372)
[25] Chen S, Sun Y, Li F, et al. Modulation of α7nAchR by Melatonin Alleviates Ischemia and Reperfusion-Compromised Integrity of Blood-Brain Barrier Through Inhibiting HMGB1-Mediated Microglia Activation and CRTC1-Mediated Neuronal Loss [published online ahead of print, 2021 Jul 1]. Cell Mol Neurobiol. 2021;10.1007/s10571-021-01122-2. doi:10.1007/s10571-021-01122-2(IF:5.046)
[26] Qian C, Al-Hamyari B, Tang X, et al. Interface-Engineered Paclitaxel-Based Hollow Mesoporous Organosilica Nanoplatforms for Photothermal-Enhanced Chemotherapy of Tumor. Mol Pharm. 2021;18(12):4531-4542. doi:10.1021/acs.molpharmaceut.1c00735(IF:4.939)
[27] Ding W, Cai C, Zhu X, Wang J, Jiang Q. Parthenolide ameliorates neurological deficits and neuroinflammation in mice with traumatic brain injury by suppressing STAT3/NF-κB and inflammasome activation. Int Immunopharmacol. 2022;108:108913. doi:10.1016/j.intimp.2022.108913(IF:4.932)
[28] Xu Y, Cao K, Guo B, et al. Lowered levels of nicotinic acetylcholine receptors and elevated apoptosis in the hippocampus of brains from patients with type 2 diabetes mellitus and db/db mice. Aging (Albany NY). 2020;12(14):14205-14218. doi:10.18632/aging.103435(IF:4.831)
[29] Wu F, Xu K, Xu K, et al. Dl-3n-butylphthalide improves traumatic brain injury recovery via inhibiting autophagy-induced blood-brain barrier disruption and cell apoptosis. J Cell Mol Med. 2020;24(2):1220-1232. doi:10.1111/jcmm.14691(IF:4.658)
[30] Xu H, Ma Z, Mo X, et al. Inducing Synergistic DNA Damage by TRIP13 and PARP1 Inhibitors Provides a Potential Treatment for Hepatocellular Carcinoma. J Cancer. 2022;13(7):2226-2237. Published 2022 Apr 11. doi:10.7150/jca.66020(IF:4.207)
[31] Wang SW, Xu Y, Weng YY, et al. Astilbin ameliorates cisplatin-induced nephrotoxicity through reducing oxidative stress and inflammation. Food Chem Toxicol. 2018;114:227-236. doi:10.1016/j.fct.2018.02.041(IF:3.977)
[32] Li Y, Xiang J, Zhang J, Lin J, Wu Y, Wang X. Inhibition of Brd4 by JQ1 Promotes Functional Recovery From Spinal Cord Injury by Activating Autophagy. Front Cell Neurosci. 2020;14:555591. Published 2020 Sep 2. doi:10.3389/fncel.2020.555591(IF:3.921)
[33] Dai L, Chen X, Xiong Y, et al. Strontium gluconate potently promotes osteoblast development and restores bone formation in glucocorticoid-induced osteoporosis rats. Biochem Biophys Res Commun. 2021;554:33-40. doi:10.1016/j.bbrc.2021.02.100(IF:3.575)
[34] Qin H, Zhang H, Zhang X, Zhang S, Zhu S, Wang H. Resveratrol attenuates radiation enteritis through the SIRT1/FOXO3a and PI3K/AKT signaling pathways. Biochem Biophys Res Commun. 2021;554:199-205. doi:10.1016/j.bbrc.2021.03.122(IF:3.575)
[35] Chen SY, Zhang GC, Shu QJ. Yang-Yin-Jie-Du decoction overcomes gefitinib resistance in non-small cell lung cancer via down-regulation of the PI3K/Akt signalling pathway. Pharm Biol. 2021;59(1):1294-1304. doi:10.1080/13880209.2021.1972122(IF:3.503)
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[40] Feng L, Liao H, Liu J, et al. Inhibition of PI3K/Akt/mTOR pathway by ammonium chloride induced apoptosis and autophagy in MAC-T cell. Res Vet Sci. 2021;136:622-630. doi:10.1016/j.rvsc.2021.01.020(IF:2.534)
[41] Wang J, Wang Y, Zuo Y, et al. MFGE8 mitigates brain injury in a rat model of SAH by maintaining vascular endothelial integrity via TIGβ5/PI3K/CXCL12 signaling. Exp Brain Res. 2021;239(7):2193-2205. doi:10.1007/s00221-021-06111-x(IF:1.972)
[42] Li L, Peng Z, Hu Q, et al. Berberine Suppressed Tumor Growth through Regulating Fatty Acid Metabolism and Triggering Cell Apoptosis via Targeting FABPs. Evid Based Complement Alternat Med. 2020;2020:6195050. Published 2020 Apr 8. doi:10.1155/2020/6195050(IF:1.813)
[43] Wu B, Liang J. Pectolinarigenin promotes functional recovery and inhibits apoptosis in rats following spinal cord injuries. Exp Ther Med. 2019;17(5):3877-3882. doi:10.3892/etm.2019.7456(IF:1.448)
[44] Wu B, Liang J. Pectolinarigenin promotes functional recovery and inhibits apoptosis in rats following spinal cord injuries. Exp Ther Med. 2019;17(5):3877-3882. doi:10.3892/etm.2019.7456(IF:1.448)
- 40307-A-TUNEL Apoptosis Detection Kit (Alexa Fluor 488)-MSDS-HB220805.pdf (PDF)
- 40307-B-TUNEL Apoptosis Detection Kit (Alexa Fluor 488)-MSDS-HB220805.pdf (PDF)
- 40307-C-TUNEL Apoptosis Detection Kit (Alexa Fluor 488)-MSDS-HB220805.pdf (PDF)
- 40307-D-TUNEL Apoptosis Detection Kit (Alexa Fluor 488)-MSDS-HB220805.pdf (PDF)
- 40307-E-TUNEL Apoptosis Detection Kit (Alexa Fluor 488)-MSDS-HB220805.pdf (PDF)
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.
