Intercalating dye with superior permeability with any live cell line
Easy no-wash mix and read protocol
Eliminates need for specialized 350nm UV laser required for Hoechst dyes and reduces chances for channel interference
No interference from small molecule fluorescence or cell autofluorescence
The NUCLEAR-ID® Green Chromatin Condensation Detection Kit detects nuclear condensation using a DNA intercalating dye that brightly stains the condensed chromatin of apoptotic cells, but only dimly stains the uncondensed chromatin of healthy living cells.
Mechanism of Action
The cell-permeable dye used in this application is an aromatic, planar cationic structure that inserts between stacked base pairs on the DNA duplex, providing an environmentally-dependent fluorescence enhancement of the dye molecules and large increases in fluorescence signal relative to the free dye in solution. Since the signal enhancement provides a proportional response, direct quantitation of DNA is possible. Further signal increase is observed upon DNA condensation during apoptosis. Considering the general mutagenic effect of nucleic acid-binding dyes, careful storage and handling of this dye is recommended.
A control apoptosis-inducing agent, staurosporine, is provided for monitoring apoptotic changes in nuclear organization. Potential applications for live-cell studies using the kit include monitoring the stages of chromatin condensation and rapid testing of compounds that induce apoptosis.
Chromatin condensation as observed by fluorescence microscopy using a standard 488nm laser. HeLa cells were treated for 4 hours with DMSO (Control) or 2 µM Staurosporine on a slide and stained with 5 µM NUCLEAR-ID® Green dye. The intercalating dye exhibits increased fluorescence upon chromatin condensation, a hallmark of apoptosis.
Flow cytometry analysis of chromatin condensation in response to application of environmentally toxic compounds. Analysis was performed following 16-hour treatment of 1 x 106 cells/ml Jurkat cells with various compounds at the indicated concentration. The 488nm excitable green-emitting dye eliminates the need for specialized UV lasers required by Hoechst dye-based assays.
Flow cytometry analysis of chromatin condensation in response to application of environmentally toxic compounds. Analysis was performed following 16-hour treatment of 1 x 106 cells/ml Jurkat cells with various compounds at the indicated concentration. The 488nm excitable green-emitting dye eliminates the need for specialized UV lasers required by Hoechst dye-based assays.
This kit provides a rapid assay for nuclear condensation, a prominent hallmark of apoptosis.
Quality Control:
Absorption peak of NUCLEAR-ID® Green dye: λmax = 507 ± 4 nm
% purity of NUCLEAR-ID® Green dye by HPLC: ≥93%
A sample from each lot of NUCLEAR-ID® Green Cell Cycle Analysis Kit is used to analyze Jurkat cells using the procedures described in the user manual. The following results are obtained:
(a) Untreated control cells: Viable cells: >85%; Apoptotic cell population: <15%
(b) Apoptosis inducer treated cells: Viable cells: >60%; Apoptotic cell population: <30%
(c) Mean fluorescence of apoptotic cells / mean fluorescence of viable cells: >40
Quantity:
200 assays
Handling:
Protect from light. Avoid freeze/thaw cycles.
Shipping:
Dry Ice
Short Term Storage:
-20°C
Long Term Storage:
-80°C
Contents:
NUCLEAR-ID® Green Cell Cycle Detection Reagent, 100 µl Apoptosis inducer (Staurosporine), 50nmoles 10X Assay Buffer, 30 ml
Scientific Background:
Apoptosis is recognized as a pathway of highly orchestrated signaling events, and of critical importance in biological processes and pathologies including development, aging and cancer. Nuclear condensation is one of the more prominent hallmarks of the many morphological features associated with apoptosis, including cell membrane blebbing, cell shrinkage, nucleosomal fragmentation, and formation of fragmented apoptotic bodies.
Regulatory Status:
RUO - Research Use Only
Product Literature References
Polyethylene glycol-functionalized poly (Lactic Acid-co-Glycolic Acid) and graphene oxide nanoparticles induce pro-inflammatory and apoptotic responses in Candida albicans-infected vaginal epithelial cells: R.D. Wagner, et al.; PLoS One 12, e0175250 (2017), Abstract; Full Text
New in vitro biomarkers to detect toxicity in human placental cells: the example of benzo[A]pyrene: A. Wakx, et al.; Toxicol. In Vitro 32, 76 (2016), Abstract;
BAG3 protects against hyperthermic stress by modulating NF-κB and ERK activities in human retinoblastoma cells: T. Yunoki, et al.; Graefes Arch. Clin. Exp. Ophthalmol. 253, 399 (2015), Abstract;
Metabolic requirements for neutrophil extracellular traps formation: O. Rodriguez-Espinosa, et al.; Immunology 145, 213 (2015), Abstract;
Genes and gene networks involved in sodium fluoride-elicited cell death accompanying endoplasmic reticulum stress in oral epithelial cells: Y. Tabuchi, et al.; Int. J. Mol. Sci. 15, 8959 (2014), Application(s): Measurement by flow cytometry, Abstract; Full Text
Inactivation of DNA-dependent protein kinase promotes heat-induced apoptosis independently of heat-shock protein induction in human cancer cell lines: S. Okazawa, et al.; PLoS One 8, e58325 (2013), Application(s): Measurement of chromatin condensation in tumor cell lines by flow cytometry, Abstract; Full Text
Inhibition of polo-like kinase 1 promotes hyperthermia sensitivity via inactivation of heat shock transcription factor 1 in human retinoblastoma cells: T. Yunoki, et al.; Invest. Ophthalmol. Vis. Sci. 54, 8353 (2013), Abstract;
Larger Daphnia at lower temperature: a role for cell size and genome configuration?: M. Jalal, et al.; Genome 56, 511 (2013), Application(s): Measurement of chromatin condensation in Daphnia nuclei using flow cytometry, Abstract; Full Text
The combination of silencing BAG3 and inhibition of the JNK pathway enhances hyperthermia sensitivity in human oral squamous cell carcinoma cells: T. Yunoki, et al.; Cancer Lett. 335, 52 (2013), Abstract;
Protective effects of veskamide, enferamide, becatamide, and oretamide on H2O2-induced apoptosis of PC-12 cells: J.B. Park; Phytomedicine 18, 843 (2011), Abstract;
Thiazole antibiotic thiostrepton synergize with bortezomib to induce apoptosis in cancer cells: B. Pandit & A.L. Gartel; PLoS One 6, (2011), Abstract; Full Text
ARC synergizes with ABT-737 to induce apoptosis in human cancer cells: U.G. Bhat, et al.; Mol. Cancer Ther. 9, 1688 (2010), Abstract;
General Literature References
Nuclear condensation and free radical scavenging: a dual mechanism of bisbenzimidazoles to modulate radiation damage to DNA.: U. Tawar et al.; Mol Cell Biochem. 305, 221 (2007), Abstract;
Three distinct stages of apoptotic nuclear condensation revealed by time-lapse imaging, biochemical and electron microscopy analysis of cell-free apoptosis.: S. Toné et al.; Exp Cell Res. 313, 3635 (2007), Abstract; Full Text
Multiparametric analysis of apoptosis by flow and image cytometry.: W. G. Telford et al.; Methods Mol. Biol. 263, 141 (2004),
Multiplex assay that distinguishes between healthy, early apoptotic, late apoptotic and necrotic cells, compatible with GFP and other fluorescent probes (blue or cyan)
Flow Cytometry, Fluorescence microscopy, Fluorescent detection | Print as PDF
Evaluating Small Molecule Cell Cycle and Chromatin Condensation Modulators Using a Novel Green Fluorescent DNA-Binding Probe: Potential Application to Compound Screening
