DNA Fragmentation Assay
For Late Apoptosis Evaluation Using the NucleoCounter® NC-3000™
- Easily measure DNA fragmentation levels
- Acquisition and analysis in one simple step
- User-friendly protocol with predefined settings
- No RNase treatment required
- No calibration required
- Clear data presentation with Plot Manager feature
- Automated PDF reports
- Export data in FCS/ACS formats
Overview of the DNA Fragmentation Assay
During apoptosis, proteins act on chromosomal DNA to fragment the DNA into ladders. This late-stage apoptotic event is detected by measuring DNA content in a way similar to that done in our Cell Cycle Assay. In apoptotic cells you will see a dissolution of peaks at the G1 and G2 phases, and the rise of a “sub G1 peak” which grows with the progression of the chromosomal DNA laddering.
The mechanism of the DNA degradation is the activation of calcium- and magnesium-dependent nucleases as apoptosis progresses. As a result, within the DNA there are nicks and double-strand breaks causing fragmentation1. This is a late-stage apoptotic event, which can be detected by quantifying the cells’ DNA content. Late-stage apoptotic cells will have less DNA content and can thereby be identified as being in the sub-G1 phase.
Assay Principle
Using fluorescence microscopy and image analysis, the NucleoCounter® NC-3000™ automatically detects cells with fragmented DNA (sub-G1 cells).
After staining fixed cells with DAPI, the sample is analyzed using the NucleoCounter® NC-3000™ system, where cellular fluorescence is quantified and apoptotic cells with fragmented DNA are identified as a sub-G1 peak in a DNA content histogram displayed on the computer screen.
Markers in the histogram can be used to demarcate apoptotic cells.
Results Presented in Plot Manager
Jurkat cells were grown in the absence (upper row) or in the presence (lower row) of camptothecin and cells were analyzed using the DNA Fragmentation Assay and a NucleoCounter® NC-3000™.
Scatter plots and histograms were obtained using the NucleoView™ NC-3000™ software. Markers in the displayed histograms were used to demarcate cells with fragmented DNA (sub-G1 cells). The colored histograms are a merge of untreated (blue line) and camptothecin treated (red line) samples.
References
- BD Larsen and CS Sørensen: The caspase‐activated DNase: apoptosis and beyond. FEBS J. 2017; 284(8): 1160-1170.
Tech notes
Publications
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166513
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
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- Instrument: NucleoCounter® NC-3000™
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- Assay: Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
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105634
- Instrument: NucleoCounter® NC-3000™
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- Assay: Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
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146:112350
- Instrument: NucleoCounter® NC-3000™
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- Assay: DNA Fragmentation Assay
- Research area: Cell Biology
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- Instrument: NucleoCounter® NC-3000™
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- Assay: DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Neurobiology
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72:105108
- Instrument: NucleoCounter® NC-3000™
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- Instrument: NucleoCounter® NC-3000™
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- Assay: Annexin V Assay, Cell Cycle Assay, Cell Vitality Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Small Molecule Drugs
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140(6):1154-1165.e5
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
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- Instrument: NucleoCounter® NC-3000™
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- Assay: Cell Cycle Assay, Cell Vitality Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Oncology
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Toxicology in vitro ,
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66:104884
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Oncology
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26(3):1465-1474
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Cell Cycle Assay, Cell Vitality Assay, DNA Fragmentation Assay
- Research area: Oncology
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- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Assay: Annexin V Assay, Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
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- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
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- Instrument: NucleoCounter® NC-3000™
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- Assay: Annexin V Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Oncology
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8(12):1505
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Assay: Annexin V Assay, Cell Count & Viability Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Cell Biology
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53:126-135
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, Cell Count & Viability Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Small Molecule Drugs
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Oncotarget ,
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2018,
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9(31):21904-21920
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
Natural antimicrobial peptides and ceragenins, as non-peptide amphipathic mimics, have been proposed as anti-cancer agents. To date, it has been confirmed that cathelicidin LL-37 and ceragenin CSA-13, both in free form and immobilized on the surface of magnetic nanoparticles (MNP@LL-37,...
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2018,
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9(5):458
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Immune Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
BRCA1-associated protein 1 (BAP1) is a nuclear deubiquitinating enzyme that is associated with multiprotein complexes that regulate key cellular pathways, including cell cycle, cellular differentiation, cell death, and the DNA damage response. In this study, we found that the reduced...
View PublicationThe Synthetic Chalcone Derivative 2-hydroxy-3′,5,5′-trimethoxychalcone Induces Unfolded Protein Response-Mediated Apoptosis in A549 Lung Cancer Cells
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Ha-Na Gil , Dongsoo Koh , Yoongho Lim , et al. ,
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Bioorganic and Medicinal Chemistry letters ,
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2018,
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28(17):2969-2975
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
The synthetic chalcone derivative 2-hydroxy-3′,5,5′-trimenthoxyochalcone (named DK-139) exhibits anti-inflammatory and anti-tumor invasion properties. However, effects of DK-139 on tumor cell growth remain unknown. In the present study, we evaluated the inhibitory activity of DK-139 against human lung cancer cells. Treatment...
View PublicationEvaluation of cytotoxicity, genotoxicity, and apoptosis of wastewater before and afterdisinfection with performic acid.
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Ragazzo P, Feretti D, Monarca S, et al. ,
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Water Res ,
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2017,
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116:44-52
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Assay: DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Small Molecule Drugs
Disinfection with performic acid (PFA) represents an emerging technology in wastewater treatment. Many recent studies indicate its effectiveness and suitability as a disinfectant for different applications; several have demonstrated its reliability as an alternative to chlorine for disinfecting secondary effluents...
View PublicationExtracellular vesicles released by fibroblasts undergoing H-Ras induced senescence show changes in lipid profile
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Buratta S, Urbanelli L, Sagini K, et al. ,
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PLoS One ,
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2017,
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12(11):e0188840
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Assay: DNA Fragmentation Assay
- Research area: Cell Biology
Cells release extracellular vesicles (EVs) in their environment and cellular lipids play an important role in their formation, secretion and uptake. Besides, there is also evidence that EV transferred lipids impact on recipient’s cell signaling. Cellular senescence is characterized by...
View PublicationBlockade of the Ras/Raf/ERK and Ras/PI3K/Akt Pathways by Monacolin K Reduces the Expression of GLO1 andInduces Apoptosis in U937 Cells
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Chen CC, Wu ML, Ho CT, Huang TC ,
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J Agric Food Chem ,
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2015,
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63(4):1186-1195
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
Deubiquitinating activity of CYLD is impaired by SUMOylation in neuroblastoma cells.
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Kobayashi T, Masoumi KC, Massoumi R. ,
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Oncogene ,
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2015,
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34(17):2251-60
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Neural Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
Differentiation and apoptosis induction by lovastatin and γ-tocotrienol in HL-60 cells via Ras/ERK/NF-κB andRas/Akt/NF-κB signaling dependent down-regulation of glyoxalase 1 and HMG-CoA reductase
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Chen CC, Liu TY, Huang SP, et al. ,
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Cell Signal ,
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2015,
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27(11):2182-90
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Assay: DNA Fragmentation Assay
- Research area: Oncology
C5-curcuminoid-4-aminoquinoline based molecular hybrids: design, synthesis and mechanistic investigation of anticancer activity
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Kandi SK, Sunny Manohar, Christian E. Vélez Gerena, et al. ,
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New J Chem ,
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2015,
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39, 224-234
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, DNA Fragmentation Assay
- Research area: Oncology
Polish natural bee honeys are anti-proliferative and anti-metastatic agents in human glioblastoma multiformeU87MG cell line.
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Moskwa J, Borawska MH, Markiewicz-Zukowska R, et al. ,
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PLoS One ,
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2014,
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9(3):e90533
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology
Xylazine as a drug of abuse and its effects on the generation of reactive species and DNA damage on humanumbilical vein endothelial cells.
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Silva-Torres L, Veléz C, Alvarez L, Zayas B. ,
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J Toxicol ,
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2014,
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492609
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Whole Blood
- Assay: DNA Fragmentation Assay
- Research area: Small Molecule Drugs
CYLD controls c-MYC expression through the JNK-dependent signaling pathway in hepatocellular carcinoma
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Pannem RR, Dorn C, Ahlqvist K, et al. ,
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Carcinogenesis. ,
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2014,
-
35(2):461-8
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Assay: Cell Count & Viability Assay, DNA Fragmentation Assay
- Research area: Oncology
Biological Activity of N-Hydroxyethyl-4-aza- 2,3-didehydropodophyllotoxin Derivatives upon Colorectal Adenocarcinoma Cells
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Vélez C, Zayas B, Kumar A. ,
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Open J Med Chem. ,
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2014,
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4(1):1-11.
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
Anticancer activity of 4-aminoquinoline-triazine based molecular hybrids
-
Manohar S, Antonella Pepe, Christian E. Vélez Gerena, et al. ,
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RSC Advances ,
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2014,
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4(14):7062-7067
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, DNA Fragmentation Assay
- Research area: Oncology
Effects of zinc supplementation and zinc chelation on in vitro β-cell function in INS-1E cells
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Nygaard SB, Larsen A, Knuhtsen A, et al. ,
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BMC Res Notes ,
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2014,
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0,35
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Immune Cells
- Assay: Cell Count & Viability Assay, Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Immunology
Imidazolium-derived ionic salts induce inhibition of cancerous cell growth through apoptosis
-
Malhotra SV, Vineet Kumar, Christian Velezb and Beatriz Zayasb ,
-
Med Chem Comm ,
-
2014,
-
5, 1404-1409
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Annexin V Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Oncology
Investigation of the cytotoxic, genotoxic, and apoptosis-inducing effects of estragole isolated from fennel(Foeniculum vulgare).
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Villarini M, Pagiotti R, Dominici L, et al. ,
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J Nat Prod ,
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2014,
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77(4):773-8
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Hepatocytes
- Assay: Cell Count & Viability Assay, Cell Cycle Assay, DNA Fragmentation Assay, Mitochondrial Potential Assay
- Research area: Cell Biology
Association of nuclear-localized Nemo-like kinase with heat-shock protein 27 inhibits apoptosis in human breastcancer cells.
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Shaw-Hallgren G, Chmielarska Masoumi K, Zarrizi R, et al. ,
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PLoS One ,
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2014,
-
9(5):e96506
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Cell Biology
Novel Nitrobenzazolo[3,2a]quinolinium Salts Induce Cell Death through a Mechanism Involving DNA Damage, Cell Cycle Changes, and Mitrochondrial Permeabilization
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Vélez C, Cox O, Rosado-Berrios CA, et al. ,
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Open J Apoptosis. ,
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2013,
-
2(2):13-22.
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: Cell Cycle Assay, DNA Fragmentation Assay
- Research area: Oncology
Expression of Id proteins is regulated by the Bcl-3 proto-oncogene in prostate cancer
-
Ahlqvist K, Saamarthy K, Syed Khaja AS, et al. ,
-
Oncogene ,
-
2013,
-
32(12):1601-8
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Mammalian Cells
- Assay: DNA Fragmentation Assay
- Research area: Immunology
Ultraviolet C Irradiation Induces Different Expression of Cyclooxygenase 2 in NIH 3T3 Cells and A431 Cells: The Roles of COX-2 Are Different in Various Cell Lines
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Tai MH, Weng CH, Mon DP, et al. ,
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Int J Mol Sci ,
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2012,
-
13(4):4351-66
- Instrument: NucleoCounter® NC-3000™
- Cell Type: Cancer Cells
- Assay: DNA Fragmentation Assay
- Research area: Oncology