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Cell Meter™ Fluorimetric Live Cell Cycle Assay Kit *Green Fluorescence Optimized for Flow Cytometry*

DNA profile in growing and nocodazole treated Jurkat cells. Jurkat cells were treated without (A) or with 100 ng/ml Nocodazole (B) in a 37 °C, 5% CO2 incubator for 24 hours, and then dye loaded with Nuclear Green™ LCS1 for 30 minutes. The fluorescence intensity of Nuclear Green™ LCS1 was measured with ACEA NovoCyte flow cytometer with the channel of FITC. In growing Jurkat cells (A), nuclear stained with Nuclear Green™ LCS1 shows G1, S, and G2 phases. In Nocodazole treated G2 arrested cells (B), frequency of G2 cells increased dramatically and G1, S phase frequency decreased significantly.
DNA profile in growing and nocodazole treated Jurkat cells. Jurkat cells were treated without (A) or with 100 ng/ml Nocodazole (B) in a 37 °C, 5% CO2 incubator for 24 hours, and then dye loaded with Nuclear Green™ LCS1 for 30 minutes. The fluorescence intensity of Nuclear Green™ LCS1 was measured with ACEA NovoCyte flow cytometer with the channel of FITC. In growing Jurkat cells (A), nuclear stained with Nuclear Green™ LCS1 shows G1, S, and G2 phases. In Nocodazole treated G2 arrested cells (B), frequency of G2 cells increased dramatically and G1, S phase frequency decreased significantly.
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Catalog Number22841
Unit Size
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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 and proliferation. There are a variety of parameters that can be used for monitoring cell viability and proliferation. In normal cells, DNA density changes depending on whether the cell is growing, dividing, resting, or performing its ordinary functions. The progression of the cell cycle is controlled by a complex interplay among various cell cycle regulators. These regulators activate transcription factors, which bind to DNA and turn on or off the production of proteins that result in cell division. Any misstep in this regulatory cascade causes abnormal cell proliferation which underlies many pathological conditions, such as tumor formation. Potential applications for live-cell studies are in the determination of cellular DNA content and cell cycle distribution for the detection of variations in growth patterns, for monitoring apoptosis, and for evaluating tumor cell behavior and suppressor gene mechanisms. This particular kit is designed to monitor cell cycle progression and proliferation using our proprietary Nuclear Green™ LCS1 in live, permeabilized and fixed cells. The percentage of cells in a given sample that are in G0/G1, S and G2/M phases, as well as the cells in the sub-G1 phase prior to apoptosis can be determined by flow cytometry. Cells stained with Nuclear Green™ LCS1 can be monitored with a flow cytometer (FL1 channel).

Platform


Flow cytometer

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

Components


Component A: 200X Nuclear Green™ LCS11 vial (250 µL)
Component B: Assay Buffer1 bottle (50 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 2.5 µL of 200X Nuclear Green™ LCS1 into 0.5 mL of cell solution
  3. Incubate at 37°C, 5% CO2 incubator for 30 - 60 minutes
  4. Analyze cells with a flow cytometer using 530/30 nm filter (FITC channel) 
Important      Thaw all the components at room temperature before starting the experiment.

CELL PREPARATION

For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. For each sample, prepare cells in 0.5 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 or other cell cycle functions.
  3. Add 2.5 µL of 200X Nuclear Green™ LCS1 (Component A) into the treated cells.
  4. Incubate the cells 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 intact, and wash the cells once with serum-containing media prior to incubation with Nuclear Green™ LCS1. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment. It is not necessary to fix the cells before DNA staining since the Nuclear Green™ LCS1 is cell-permeable.
  5. Optional: Centrifuge the cells at 1000 rpm for 4 minutes, and then re-suspend cells in 0.5 mL of assay buffer (Component B) or the buffer of your choice.
  6. Monitor the fluorescence intensity using a flow cytometer using 530/30 nm filter (FITC channel). Gate on the cells of interest, excluding debris. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)503
Emission (nm)527

Citations


View all 1 citations: Citation Explorer
Antineoplastic Effects and Mechanisms of a New RGD Chimeric Peptide from Bullfrog Skin on the Proliferation and Apoptosis of B16F10 Cells
Authors: Jiang, Xuan and Zhang, Xin and Fu, Chao and Zhao, Ruili and Jin, Tianming and Liu, Mengyue and Pan, Chenhao and Li, Liu An and Ma, Jifei and Yu, Enyuan and others,
Journal: The Protein Journal (2021): 1--12

References


View all 33 references: Citation Explorer
Cell cycle synchronization of Escherichia coli using the stringent response, with fluorescence labeling assays for DNA content and replication
Authors: Ferullo DJ, Cooper DL, Moore HR, Lovett ST.
Journal: Methods (2009): 8
DNA replication, cell cycle progression and the targeted gene repair reaction
Authors: Engstrom JU, Kmiec EB.
Journal: Cell Cycle (2008): 1402
Morin inhibits the growth of human leukemia HL-60 cells via cell cycle arrest and induction of apoptosis through mitochondria dependent pathway
Authors: Kuo HM, Chang LS, Lin YL, Lu HF, Yang JS, Lee JH, Chung JG.
Journal: Anticancer Res (2007): 395
Direct control of cell cycle gene expression by proto-oncogene product ACTR, and its autoregulation underlies its transforming activity
Authors: Louie MC, Revenko AS, Zou JX, Yao J, Chen HW.
Journal: Mol Cell Biol (2006): 3810
Cell cycle markers for live cell analyses
Authors: Easwaran HP, Leonhardt H, Cardoso MC.
Journal: Cell Cycle (2005): 453
Dynamic relocalization of hOGG1 during the cell cycle is disrupted in cells harbouring the hOGG1-Cys326 polymorphic variant
Authors: Luna L, Rolseth V, Hildrestr and GA, Otterlei M, Dantzer F, Bjoras M, Seeberg E.
Journal: Nucleic Acids Res (2005): 1813
Dynamics of relative chromosome position during the cell cycle
Authors: Essers J, van Cappellen WA, Theil AF, van Drunen E, Jaspers NG, Hoeijmakers JH, Wyman C, Vermeulen W, Kanaar R.
Journal: Mol Biol Cell (2005): 769
Description of a flow cytometry approach based on SYBR-14 staining for the assessment of DNA content, cell cycle analysis, and sorting of living normal and neoplastic cells
Authors: Nunez R, Garay N, Villafane C, Bruno A, Lindgren V.
Journal: Exp Mol Pathol (2004): 29
Videomicrofluorometry on living cells and discriminant factorial analysis to study cell cycle distributions
Authors: Savatier J, Gbankoto A, Vigo J, Salmon JM.
Journal: J Biol Regul Homeost Agents (2004): 206
Cell cycle regulation of the murine 8-oxoguanine DNA glycosylase (mOGG1): mOGG1 associates with microtubules during interphase and mitosis
Authors: Conlon KA, Zharkov DO, Berrios M.
Journal: DNA Repair (Amst) (2004): 1601