For dose-response proliferation assays, 4000 cells were seeded in 96-well culture plates in complete medium. After 24 hours, the indicated concentrations of sotorasib and cetuximab were added to cell. After 72 hours, cell viability was determined by measuring ATP content using Cell Titer-Glo® Luminescent Cell Viability assay (Promega). DMSO-only treated cells were used as control. Assays were performed with 3 replicates and were each repeated three times. For senolytic proliferation assays with RW7213-R, 2000 cells were seeded in 96-well culture plates in medium containing cetuximab 50μg/mL and sotorasib 3μM combo or medium only to induce senescence. After 96 hours, serial dilutions of AZD8055 or navitoclax were added to cells and cell viability was determined at baseline by measuring ATP content using Cell Titer-Glo® Luminescent Cell Viability assay (Promega). After 96 hours cell viability was determined again by measuring ATP content using Cell Titer-Glo® Luminescent Cell Viability assay (Promega). DMSO-only treated cells were used as control and all the values were normalized on baseline measurements. In all the experiments, plates were incubated at 37°C in 5% CO2.
Celltiter glo luminescent cell viability assay
Manufactured by Promega
Sourced in United States, Germany, United Kingdom, Switzerland, Italy, France, Japan, Spain, Sweden, Belgium, China, Australia
The CellTiter-Glo Luminescent Cell Viability Assay is a quantitative method for determining the number of viable cells in a cell-based assay. The assay measures the amount of ATP present, which is an indicator of metabolically active cells.
Automatically generated - may contain errors
Lab products found in correlation
Caspase glo 3 7 assay, by Promega (63 mentions)
Glomax 96 microplate luminometer, by Promega (31 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (31 mentions)
Glomax multi detection system, by Promega (26 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (23 mentions)
Prism 7, by GraphPad (23 mentions)
Enspire multimode plate reader, by PerkinElmer (22 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (22 mentions)
Celltiter glo reagent, by Promega (21 mentions)
96 well plate, by Corning (21 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (19 mentions)
Calcusyn software, by Biosoft (17 mentions)
Infinite 200 pro, by Tecan (17 mentions)
Prism software, by GraphPad (17 mentions)
Prism 8, by GraphPad (17 mentions)
Prism 6, by GraphPad (17 mentions)
Fluostar omega, by BMG Labtech (14 mentions)
Celltiter 96 aqueous one solution cell proliferation assay, by Promega (14 mentions)
Glomax luminometer, by Promega (14 mentions)
Lactate assay kit, by Abcam (14 mentions)
2 982 protocols using celltiter glo luminescent cell viability assay
1
Dose-Response and Senolytic Assays
2
TNFα Cytotoxicity Assay in Jurkat Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Parental Jurkat E6-1 cells or TNFR2 overexpressing Jurkat single clone cells were plated in opaque 96 well plates with 104 cells in 100μL complete RPMI medium per well. Gradient concentration of human TNFα (Gibco, PHC3011) was added. The cell viability after 24 hours culture was measured with Promega CellTiter-Glo® Luminescent Cell Viability Assay (Promega, G7570) according to manufacturer’s instructions. AN3025 or human IgG1, κ (BioxCell, BE0297) was added to the hTNFR2 overexpressing Jurkat cells by dose titration along with constant human TNFα (0.5 ng/ml). The cell viability after 24 hours culture was measured with Promega CellTiter-Glo® Luminescent Cell Viability Assay (Promega, G7570).
3
Thorax Homogenization and ATP/Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thoraxes from the 20 flies were homogenized in CellTiter-Glo® buffer [Celltiter-Glo® Luminescent Cell Viability Assay (Promega))], and ATP was measured by using the Celltiter-Glo® Luminescent Cell Viability Assay (Promega) according to the manufacturer’s manual. Protein levels were measured with the BCA Protein Assay kit (Thermo Fisher Scientific).
4
Comparative Cell Viability Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mock/shRNASET2-OAW42 cells were seeded at 3000 cells/well in 96-well plates. At the end of each time point, mitochondrial activity was measured with CellTiter-Glo® Luminescent Cell Viability Assay, accordingly to the manufacturer’s instructions (Promega, Madison, WI, USA). Mock/ tRNASET2-SKOV3 cells were seeded at 10,000 cells/well in 48-well plates. At the end of each time point, cells were trypsinized, resuspended in medium and immediately counted. We did not analyze mock and tRNASET2-SKOV3 cells’ mitochondrial activity, as described for OAW42, due to interference of GFP protein expressed from cells and the assay. For 3D culture, cells were grown in AlgimatrixTM (ThermoFisher Scientific) for 10 days and then recovered after AlgimatrixTM dissolving, according to the manufacturers’ protocols. Each well was divided form manual count of the spheres and for evaluation of cell viability with CellTiter-Glo® Luminescent Cell Viability Assay (Promega). Cell viability assay was performed with PP2 (0, 2.5, 5, 10, 20, and 40 μM) up to 72 h. Mock/shRNASET2-OAW42 cells were seeded at 3000 cells/well in a 96-well plate; then cell viability was assessed with CellTiter-Glo® Luminescent Cell Viability Assay. Mock/tRNASET2-SKOV3 cells were seeded at 25,000 cells/well in a 24-well plate; at 72 h cells were trypsinized, resuspended in medium, and immediately counted.
5
Inhibition of Cancer Cell Growth
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 11
The ability of a compound of the present disclosure to inhibit the growth of cells, such as human leukemia, VCaP, LNCaP, 22RV1, DU145, LNCaP-AR, MV4;11, KOPN-8, ML-2, MOLM-13, RS4;11, SEM, bone marrow cells (BMCs), MLL-AF9, MLL-AF4, MLL-ENL, MLL-CBP, MLL-GAS7, MLL-AF1p, MLL-AF6, HM-2, E2A-HLF, REH, U937, K562, KG-1, HL-60 and NB4 cells, is tested using a cell viability assay, such as the Promega CellTiter-Glo® Luminescent Cell Viability Assay (Promega Technical Bulletin, 2015, “CellTiter-Glo® Luminescent Cell Viability Assay”: 1-15, herein incorporated by reference in its entirety). Cells are plated at relevant concentrations, for example about 1×105-2×105 cells per well in a 96-well plate. A compound of the present disclosure is added at a concentration up to about 2 μM with eight, 2-fold serial dilutions for each compound. Cells are incubated at 37° C. for a period of time, for example, 72 hours, then cells in the control wells are counted. Media is changed to restore viable cell numbers to the original concentration, and compounds are re-supplied. Proliferation is measured about 72 hours later using Promega CellTiter-Glo® reagents, as per kit instructions.
6
Cell proliferation and drug synergy assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the effects of compounds on cell proliferation, cells were plated in 96-well plates at a density of 5000–10,000 cells per well to achieve 70% of confluence in a volume of 100 µL, and grown for 24 hr prior to treatment. Cells were then treated with DMSO or compounds at concentrations ranging from 1.5 nM to 10 µM (threefold dilutions). After 6 days, cell proliferation was measured using the CellTiter-Glo luminescent cell viability assay (Promega, Madison, WI) and Victor4 plate reader (Perkin Elmer, Waltham, MA). Percent inhibition was calculated relative to DMSO signal, and all results shown are the mean of triplicate measurements.
For synergy analyses of drug combinations, 5000–10,000 cells suspended in 100 µl media were plated in triplicate into each well of 96-well tissue culture plates and grown for 24 hr prior to treatments. Ceritinib and CGM097 at different concentrations defined by a dose matrix were added to the cells such that all pair-wise combinations as well as the single agents were represented. Cells were incubated for 72 hr following compound addition, and cell viability was measured using the CellTiter-Glo luminescent cell viability assay (Promega) and Victor4 plate reader (Perkin Elmer). Isobolograms and combination indices were determined as described by Lehar et al. (Lehár et al., 2009 (link)).
For synergy analyses of drug combinations, 5000–10,000 cells suspended in 100 µl media were plated in triplicate into each well of 96-well tissue culture plates and grown for 24 hr prior to treatments. Ceritinib and CGM097 at different concentrations defined by a dose matrix were added to the cells such that all pair-wise combinations as well as the single agents were represented. Cells were incubated for 72 hr following compound addition, and cell viability was measured using the CellTiter-Glo luminescent cell viability assay (Promega) and Victor4 plate reader (Perkin Elmer). Isobolograms and combination indices were determined as described by Lehar et al. (Lehár et al., 2009 (link)).
7
Cell Viability Assay in Endothelial, Fibroblast, and Carcinoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 2
HUVEC (human umbilical vein endothelial cells), NHDF (normal human dermal fibroblasts) and A549 (human epithelial-like carcinoma cells) cells were seeded at three different cell densities into 96-well plates and were treated with Aposec (2.5 U/mL), bFGF (20 ng/mL) and TNFa (4 ng/mL). In addition to the cell numbers and treatments, the cells were cultivated with different percentages of growth medium. After an overnight incubation in the CO2-incubator, the cell number was determined using CellTiter-Glo Luminescent Cell Viability As-say (CellTiterGlo), which is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present (it is an indicator of metabolically active cells) (see Technical Bulletin, CellTiter-Glo Luminescent Cell Viability Assay, Promega, TB288, Revised 3/15).
A clear cell density dependent cell number was observed upon the overnight incubation. No sign for a proliferation induction by Aposec could be observed (see
8
Cell Viability Assay for Anti-Cancer Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 11
The ability of a compound of the present disclosure to inhibit the growth of cells, such as human leukemia, VCaP, LNCaP, 22RV1, DU145, LNCaP-AR, MV4;11, KOPN-8, ML-2, MOLM-13, RS4;11, SEM, bone marrow cells (BMCs), MLL-AF9, MLL-AF4, MLL-ENL, MLL-CBP, MLL-GAS7, MLL-AF1p, MLL-AF6, HM-2, E2A-HLF, REH, U937, K562, KG-1, HL-60 and NB4 cells, is tested using a cell viability assay, such as the Promega CellTiter-Glo® Luminescent Cell Viability Assay (Promega Technical Bulletin, 2015, “CellTiter-Glo® Luminescent Cell Viability Assay”: 1-15, herein incorporated by reference in its entirety). Cells are plated at relevant concentrations, for example about 1×105-2×105 cells per well in a 96-well plate. A compound of the present disclosure is added at a concentration up to about 2 μM with eight, 2-fold serial dilutions for each compound. Cells are incubated at 37° C. for a period of time, for example, 72 hours, then cells in the control wells are counted. Media is changed to restore viable cell numbers to the original concentration, and compounds are re-supplied. Proliferation is measured about 72 hours later using Promega CellTiter-Glo® reagents, as per kit instructions.
9
Cell Viability Assay Protocols
10
Evaluating Compound Cytotoxicity and Synergy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were plated in 96-well plates at 2000 cells per well, 24 hours before treatment. Cells were treated with serial dilutions of indicated compounds alone or in the presence of 3 μM LIMKi or DMSO, for 72 hours. Surviving cells were fixed with 4% paraformaldehyde and stained with 250 ng/ml DAPI. Experiments were repeated with three independent replicate experiments. Nuclei were imaged on a High Content Imaging Operetta system (PerkinElmer) and quantified using Harmony® High Content Imaging and Analysis Software (PerkinElmer). Cell numbers were plotted as percent change from DMSO-treated control and EC50 values were calculated from dose-response curves using Prism 5 (GraphPad). Drug combination synergy was determined by treating cells with serial dilutions of LIMKi (2.5-20 μM) and Vincristine (0.625-5 nM) alone or in 4×4 combinations, and quantifying cells 72 hours using CellTiter-Glo® Luminescent Cell Viability Assay (Promega), following manufacturer's protocol. Combination index and effect parameters were determined with CalcuSyn [36 (link)]. Neuroblastoma cell viability was quantified using CellTiter-Glo® Luminescent Cell Viability Assay (Promega), following manufacturer's protocol.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!