Cyquant proliferation kit

Manufactured by Thermo Fisher Scientific

The CyQUANT proliferation kit is a fluorescence-based assay used to quantify cell proliferation in cell culture experiments. The kit measures the total cellular nucleic acid content, which is directly proportional to the number of cells present. This provides an objective and accurate method for determining cell growth and proliferation.

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Lab products found in correlation

Incucyte, by Sartorius (1 mentions) Hema3 kit, by Thermo Fisher Scientific (1 mentions)

8 protocols using cyquant proliferation kit

Measurement of DNA Synthesis in VVEC

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To determine the rate of DNA synthesis, VVEC were plated in 24-well plates at a density of 1.2 × 10⁴ cells/well in DMEM supplemented with 10% FBS. On the next day, cells were rinsed with phosphate-buffered saline (PBS), and incubated in DMEM without serum for 72 h. Cells were preincubated with 2-deoxyglucose (2-DG; 2 mM), oligomycin (100 ng/ml), rotenone (0.1 μM), or FCCP (5 μM) for 20 min and stimulated with ATP, 2-methylthioadenosine diphosphate trisodium salt (MeSADP), or adenosine (100 μM each) in the presence of 0.125 μCi of [methyl-³H]thymidine (NEN Life Science Products, Boston, MA) for 24 h. Incorporated radioactivity was measured using a β-counter, as previously described (30 (link)). To determine the effects of metabolic inhibitors on VVEC proliferation, cells were plated in 96-well plates at a density of 1.2 × 10³ cells/well in DMEM supplemented with 1% FBS and grew in the presence of indicated metabolic inhibitors. Incubation media with all indicated components were changed each second day, and cell proliferation rate was assessed using CyQuant proliferation kit (Invitrogen, Carlsbad, CA), according to the manufacturer's instructions. Fluorescence intensity was determined using a plate reader (BMG Labtech).

Lapel M., Weston P., Strassheim D., Karoor V., Burns N., Lyubchenko T., Paucek P., Stenmark K.R, & Gerasimovskaya E.V. (2016). Glycolysis and oxidative phosphorylation are essential for purinergic receptor-mediated angiogenic responses in vasa vasorum endothelial cells. American Journal of Physiology - Cell Physiology, 312(1), C56-C70.

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Cell Proliferation Assay Protocol

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Cells were plated in 24 or 96 well trays, reverse-transfected then scanned over 72 h by Incucyte (Essen BioScience). For proliferation assays, cells were reverse-transfected then cell numbers measured over various time points using either a CyQUANT proliferation kit (Invitrogen) or CellTiter 96 Aqueous MTS proliferation assay kit (Promega) according to manufacturer’s instructions.

Yu F., Pillman K.A., Neilsen C.T., Toubia J., Lawrence D.M., Tsykin A., Gantier M.P., Callen D.F., Goodall G.J, & Bracken C.P. (2017). Naturally existing isoforms of miR-222 have distinct functions. Nucleic Acids Research, 45(19), 11371-11385.

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Cell Proliferation Quantification Assay

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Cell proliferation analysis was performed as described previously (15 (link)). Briefly, 2000 cells/well were plated in 96 well plates in triplicate, followed by different treatments for the indicated time. The cell proliferation rate was assessed using CyQUANT proliferation kit (Invitrogen) following the manufacturer’s instructions. Absorbance at 480 and 650 nm was measured using a TECAN plate reader. The relative cell proliferation rate at each time point was determined relative to time zero.

Cairns J., Ingle J.N., Kalari K.R., Goetz M.P., Weinshilboum R.M., Gao H., Li H., Bari M.G, & Wang L. (2021). Anastrozole regulates fatty acid synthase in breast cancer. Molecular cancer therapeutics, 21(1), 206-216.

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Transwell Boyden Chamber Invasion Assay

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Cell invasion was evaluated using the Transwell Boyden chamber system, following the protocol described by New et al. in 2017. In brief, 8-μm pore inserts were utilized for migration assessment. A thin layer of VitroGel was applied to the inserts. HNSCC cells (3 × 10⁴ cells/well) within serum-free culture media were plated on the VitroGel-coated inserts. These inserts were then set in triplicate containment wells containing media containing serum up to 24 hours. Simultaneously, cells (2 × 10³ cells/well in a 96-well plate) were plated under identical conditions to assess viability using the CyQUANT assay. Following fixative treatment then staining with the Hema 3 kit (Thermo Fisher Scientific), quantity of cells that invaded through the VitroGel and across the Boyden chamber membrane was quantified. The number of invading cells was normalized based on cell viability. To measure viability, HNSCC cells were seeded in quadruplicate (2 × 10³ cells/well in a 96-well plate) at initial day 0. Experimental conditions were applied on day 1 for durations of 24 to 72 hours, and cellular viability was quantified using a CyQUANT proliferation kit (Invitrogen C7026) pursuant to the manufacturer’s guidelines.

Arnold L., Yap M., Jackson L., Barry M., Ly T., Morrison A., Gomez J.P., Washburn M.P., Standing D., Yellapu N.K., Li L., Umar S., Anant S, & Thomas S.M. (2024). DCLK1-Mediated Regulation of Invadopodia Dynamics and Matrix Metalloproteinase Trafficking Drives Invasive Progression in Head and Neck Squamous Cell Carcinoma. bioRxiv.

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Fibroblast Proliferation Assay

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Fibroblasts/ml (5 × 10⁴) in a total volume of 100 µl were seeded in a 96-well plate, and proliferation was determined using the CyQuant proliferation kit (Invitrogen) as per the manufacturers’ protocol.

Ahadome S.D., Mathew R., Reyes N.J., Mettu P.S., Cousins S.W., Calder V.L, & Saban D.R. (2016). Classical dendritic cells mediate fibrosis directly via the retinoic acid pathway in severe eye allergy. JCI Insight, 1(12), e87012.

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Cell Cytotoxicity Assay with TAF and Ficlatuzumab

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The HN5 and UM-SCC-1 cells were seeded in 96-well plates (1500 cells per well), and after 12 hours cells were treated with DMEM alone or TAF-conditioned media with or without ficlatuzumab at various doses. After 72 hours of treatment, media were discarded and cells subjected to a freeze-thaw cycle at −80°C for 10 minutes followed by 37°C for 10 minutes to completely lyse the cells. Cell viability was determined using the CyQuant proliferation kit (Life Technologies) according to the manufacturer’s instructions.

Kumar D., Kandl C., Hamilton C.D., Shnayder Y., Tsue T.T., Kakarala K., Ledgerwood L., Sun X.S., Huang H.(., Girod D, & Thomas S.M. (2015). Mitigation of Tumor-Associated Fibroblast-Facilitated Head and Neck Cancer Progression With Anti–Hepatocyte Growth Factor Antibody Ficlatuzumab. JAMA otolaryngology-- head & neck surgery, 141(12), 1133-1139.

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CyQuant Cell Proliferation Assay

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Cell number was measured using the CyQuant proliferation kit (Life Technologies, Grand Island, NY) according to the manufacturer’s protocol. Briefly, cells were plated (2,000 cells/well) in 96 well plates and assayed 72 h later following various experimental treatment conditions in serum free media. Cells were lysed at −80° C, and CyQuant dye used to measure relative cell number by fluorescence assessment.

Kumar D., Yalamanchali S., New J., Parsel S., New N., Holcomb A., Gunewardena S., Tawfik O., Lominska C., Kimler B.F., Anant S., Kakarala K., Tsue T.T., Shnayder Y., Sykes K., Padhye S, & Thomas S.M. (2018). Development and characterization of an in vitro model for radiation-induced fibrosis. Radiation research, 189(3), 326-336.

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Cell Viability Assay and Radiation Exposure

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HNSCC cells were seeded in triplicate (2000 cells/well, 96-well plate). After cells had adhered, various experimental conditions were applied for 72 h duration. Cell viability was assessed using CyQuant proliferation kit (life technologies) according to manufacturer’s instructions. For irradiation experiments, plates were exposed to gamma radiation (J.L. Shepherd and Associates Mark I Model 68A cesium-137 source irradiator; dose rate = 2.9 Gy/min).

New J., Arnold L., Ananth M., Alvi S., Thornton M., Werner L., Tawfik O., Dai H., Shnayder Y., Kakarala K., Tsue T.T., Girod D., Ding W.X., Anant S, & Thomas S.M. (2017). Secretory autophagy in cancer-associated fibroblasts promotes head and neck cancer progression and offers a novel therapeutic target. Cancer research, 77(23), 6679-6691.

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