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Cell Meter™ Nuclear Apoptosis Assay Kit *Green Fluorescence Optimized for Flow Cytometry*

The increase in fluorescence intensity of Nuclear Green™ DCS1 with the addition of Camptothecin in Jurkat cells. Jurkat cells were treated overnight without (Left) or with 20 μM camptothecin (Right) in a 37 oC, 5% CO2 incubator, and then dye loaded with Nuclear Green™ DCS1 for 60 minutes. At the end of 15 minutes of Nuclear Green™ DCS1 dye loading, MitoLite™ NIR (Cat. # 22802) was added for multicolor analysis. The fluorescence intensity of Nuclear Green™ DCS1 and MitoLite™ NIR was measured with a FACSCalibur (Becton Dickinson, San Jose, CA) flow cytometer using FL1 channel (Nuclear Green™ DCS1) and FL4 channel (MitoLite™ NIR).
The increase in fluorescence intensity of Nuclear Green™ DCS1 with the addition of Camptothecin in Jurkat cells. Jurkat cells were treated overnight without (Left) or with 20 μM camptothecin (Right) in a 37 oC, 5% CO2 incubator, and then dye loaded with Nuclear Green™ DCS1 for 60 minutes. At the end of 15 minutes of Nuclear Green™ DCS1 dye loading, MitoLite™ NIR (Cat. # 22802) was added for multicolor analysis. The fluorescence intensity of Nuclear Green™ DCS1 and MitoLite™ NIR was measured with a FACSCalibur (Becton Dickinson, San Jose, CA) flow cytometer using FL1 channel (Nuclear Green™ DCS1) and FL4 channel (MitoLite™ NIR).
Ordering information
Price ()
Catalog Number22811
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)503
Emission (nm)527
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
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OverviewpdfSDSpdfProtocol


Excitation (nm)
503
Emission (nm)
527
Our Cell Meter™ assay kits are a set of tools for monitoring cell viability. There are a variety of parameters that can be used for monitoring cell viability. This particular kit is designed to monitor cell apoptosis through measuring the apoptotic chromatin condensation. The compacted chromatin of apoptotic cells binds higher amounts of nuclear dye compared to the healthy cells. Our Cell Meter™ Nuclear Apoptosis Assay Kit provides all the essential components with an optimized assay method for the detection of apoptosis in cells with condensed nuclei. This fluorometric assay is based on the detection of the DNA contents in cells using our proprietary non-fluorescent dye that becomes strongly fluorescent upon binding to DNA. In normal cells, Nuclear Green™ is not cell permeable, however, in apoptotic cells, Cells with compromised plasma membranes or with impaired/no cell metabolism are unable to prevent the dye from entering the cell. Once inside the cell, the dyes bind to intracellular DNA producing highly fluorescent complexes which identify the cells as non-viable cells. The staining with Nuclear Green; DCS1 can be measured using a flow cytometer (FL1 channel) or fluorescence microscope (FITC filter set). The kit can be used with our other apoptosis reagents, such as Our Cell Meter™ NIR Mitochondria Membrane Potential Detection Kit (Cat# 22802), for multi-parametric study of cell viability and apoptosis. The kit is optimized for screening of apoptosis activators and inhibitors.

Platform


Flow cytometer

Excitation488 nm laser
Emission530/30 nm filter
Instrument specification(s)FITC channel

Components


Component A: 200X Nuclear Green™ DCS11 vial (500 µL)
Component B: Assay Buffer1 bottle (100 mL)

Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cells with test compounds at a density of 5 × 105 to 1 × 106 cells/mL
  2. Add 5 µL of 200X Nuclear Green™ DCS1 into 1 mL of cell solution
  3. Incubate the cells in a 37°C, 5% CO2 incubator for 30 to 60 minutes
  4. Pellet the cells and resuspend the cells in 1 mL of growth medium
  5. Analyze the fluorescence intensity using flow cytometer with FL1 channel

Important notes
Thaw all the kit components at room temperature before starting the experiment.

SAMPLE EXPERIMENTAL PROTOCOL

  1. For each sample, prepare cells in 1 mL of warm medium or buffer of your choice at a density of 5 × 105 to 1 × 106 cells/mL. Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for apoptosis induction.

  2. Treat cells with test compounds for a desired period of time to induce apoptosis.

  3. Add 5 µL of 200X Nuclear Green™ DCS1 (Component A) into the treated cells.

  4. Incubate the cell solution in a 37°C, 5% CO2 incubator for 30 to 60 minutes. Note: For adherent cells, gently lift the cells with 0.5 mM EDTA to keep the cells intack, and wash the cells once with serum-containing media prior to the incubation with Nuclear Green™ DCS1 dye-loading solution. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.

  5. Optional: Centrifuge the cells at 1000 rpm for 4 minutes, and then re-suspend cells in 1 mL of Assay Buffer (Component B) or buffer of your choice.

  6. Monitor the fluorescence intensity using a flow cytometer with FL1 channel (Ex/Em = 490/525 nm). Gate on the cells of interest, excluding debris.

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)503
Emission (nm)527

References


View all 93 references: Citation Explorer
Curcumin induces cell cycle arrest and apoptosis in human osteosarcoma (HOS) cells
Authors: Lee DS, Lee MK, Kim JH.
Journal: Anticancer Res (2009): 5039
Real-time in vivo imaging of retinal cell apoptosis after laser exposure
Authors: Schmitz-Valckenberg S, Guo L, Maass A, Cheung W, Vugler A, Moss SE, Munro PM, Fitzke FW, Cordeiro MF.
Journal: Invest Ophthalmol Vis Sci (2008): 2773
Morphological study on apoptosis Hela cells induced by soyasaponins
Authors: Xiao JX, Huang GQ, Zhu CP, Ren DD, Zhang SH.
Journal: Toxicol In Vitro (2007): 820
Effects of neuraminidase on apoptosis of blood lymphocytes in rats with implanted Morris tumor
Authors: Grata-Borkowska U, Steciwko A, Pokorski M, Drobnik J, Gasiorowski K, Pirogowicz I, Cieslar-Marczak E.
Journal: J Physiol Pharmacol (2007): 253
Effects of Chinese herbal recipe Weichang'an in inducing apoptosis and related gene expression in human gastric cancer grafted onto nude mice
Authors: Zhao AG, Yang JK, You SF, Li T, Zhao HL, Gu Y, Tang LD, Qiu JX.
Journal: Zhong Xi Yi Jie He Xue Bao (2007): 287
Growth arrest and apoptosis induced by quercetin is not linked to adipogenic conversion of human preadipocytes
Authors: Morikawa K, Ikeda C, Nonaka M, Suzuki I.
Journal: Metabolism (2007): 1656
Severe acute respiratory syndrome coronavirus 3C-like protease-induced apoptosis
Authors: Lin CW, Lin KH, Hsieh TH, Shiu SY, Li JY.
Journal: FEMS Immunol Med Microbiol (2006): 375
Apoptosis induction and mitochondria alteration in human HeLa tumour cells by photoproducts of Rose Bengal acetate
Authors: Panzarini E, Tenuzzo B, Palazzo F, Chionna A, Dini L.
Journal: J Photochem Photobiol B (2006): 39
Cytoplasmic Ca2+ signals and cellular death by apoptosis in myocardiac H9c2 cells
Authors: Lax A, Soler F, Fern and ez-Belda F., undefined
Journal: Biochim Biophys Acta (2006): 937
Hydroxycamptothecin-induced apoptosis in hepatoma SMMC-7721 cells and the role of mitochondrial pathway
Authors: Fu YR, Yi ZJ, Yan YR, Qiu ZY.
Journal: Mitochondrion (2006): 211