Cancer cell lines (one cell line for each organ represented in the NCI-60 panel) and normal cell lines (3000-5000/well) were seeded in 96 well microplates (white plates from Nunc to block luminescence bleeding between the wells), and allowed to attach for 24 h. Then cells were incubated for 3 h with peptides (50μΜ of 9R, 9S1R and 124R), or 100 mM sodium azide as a control. After 3 h, 10 μl of a single reagent from Cell Titer Glow™ (Promega) was added to cells. For the cellular supernatant ATP content, the supernatant of each well was transferred to a new microplate and 10 μl of a single reagent from Cell Titer Glow™ (Promega) was added to each supernatant. Complete reagent mixing in 96 wells plates required gentle orbital shaking for 2–10 min. The plate reading was taken by a SynergyMx plate reader. Plate readings were exported to Microsoft Excel and Graph-PadPrism software. All the wells were analyzed in triplicates. The statistical analysis was done with the GraphPadPRISM*.
Cell titer glow
Manufactured by Promega
Sourced in United States
Cell Titer Glow is a luminescent cell viability assay that quantifies the number of viable cells in a sample. 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
Selumetinib, by Selleck Chemicals (3 mentions)
Lxs196, by MedChemExpress (3 mentions)
Facsdiva 6, by BD (2 mentions)
96 well microtiter plate luminometer, by Dynex (2 mentions)
Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Spectramax id3 multi mode microplate reader, by Molecular Devices (1 mentions)
Ganciclovir, by Merck Group (1 mentions)
Incucyte s3 live cell analysis system, by Sartorius (1 mentions)
Pnd 1186, by Selleck Chemicals (1 mentions)
Vs 6063, by Selleck Chemicals (1 mentions)
G7570, by Promega (1 mentions)
Envision plate reader, by PerkinElmer (1 mentions)
Celltiter glo cell viability assay kit, by Promega (1 mentions)
Glowmax, by Promega (1 mentions)
D300e digital, by Tecan (1 mentions)
H9 derived, by Thermo Fisher Scientific (1 mentions)
Nhdf ad, by Lonza (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
96 well plate reader, by Tecan (1 mentions)
Doxorubicin, by MedChemExpress (1 mentions)
37 protocols using cell titer glow
1
ATP Content Assay for Cell Viability
2
Cell Viability and Proliferation Assays
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Cell viability was analyzed using the AlamarBlue cell viability reagent (Thermo Fisher) or CellTiter-Glow (Promega) according to the manual. Fluorescence emission was recorded with a SpectraMax iD3 Multi-Mode Microplate Reader (Molecular Devices). To determine whether introduced mutations into the HBEGF locus affect the proliferative capacity, we evaluated cell growth of HEK293 wild-type cells and the produced HBEGF-mutant sublines. To monitor proliferation curves, 2000 cells were seeded per well of a 96-well plate and cell confluence was recorded every 24 h for 7 days, using the Incucyte S3 live-cell analysis system (Essen BioScience). For the experiments with the HSV-TK safety switch in hiPSCs, ganciclovir (Sigma, SML2346) was included in the cell culture media at 0.1, 1, and 10 µM concentration for 3 days, followed by a 3-day recovery.
3
Evaluating FAK Inhibition on Cell Viability and Invasion
4
Comparative Analysis of Uveal Melanoma Cell Inhibition
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Example 90
Procedure: Uveal melanoma cell lines, 92-1 or MP41, were plated in 96 well plates in the presence of growth media (See Table 7). BAF ATPase inhibitors (Compound 87), PKC inhibitor (LXS196; MedChemExpress), or MEK inhibitor (Selumetinib; Selleck Chemicals) were dissolved in DMSO and added to the cells in a concentration gradient from 0 to 10 micromolar at the time of plating. Cells were incubated at 37 degrees Celsius for 3 days. After three days of treatment, cell growth was measured with Cell-titer glow (Promega), and luminescence was read on an Envision plate reader (Perkin Elmer).
Results: As shown in
5
Inhibition of Cell Growth by BRG1/BRM ATPase Inhibitor
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Example 12
Procedure: Uveal melanoma cell lines (92-1, MP41, MP38, MP46) and non-small cell lung cancer cells (NCIH1299) were plated into 96 well plates with growth media (see Table 2). BRG1/BRM ATPase inhibitor, Compound B, was dissolved in DMSO and added to the cells in a concentration gradient from 0 to 10 μM at the time of plating. Cells were incubated at 37° C. for 3 days. After three days of treatment, cell growth was measured with Cell-titer glow (Promega), and luminescence was read on an Envision plate reader (Perkin Elmer).
Results: As shown in
6
Quantifying Neurosphere Formation Potential
7
Cell Viability and Drug Sensitivity Assay
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Cell viability and drug sensitivity were measured using a CellTiter-Glo® Cell Viability Assay Kit (Promega) according to the manufacturer’s protocol. Briefly, cells were cultured in sextuplicate in 96-well plates (∼250 cells per well) and incubated for 24 h to allow cell attachment on the surface of the wells in charcoal-stripped serum. Then, cells were exposed to different concentration of drugs. The effects on cell viability were tested when drugs present in the culture medium. On day six, cell viability was assessed by adding 100 μL per well of cell titer glow (Promega) followed by a 10 min incubation at 37 °C and measured by the amount of luminescence using a 96-well plate luminometer (GlowMax; Promega). Background was subtracted using the medium-only control wells.
8
High-Throughput Synergistic Drug Screening
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Cells were plated in white-walled 384-well plates (Greiner) at predetermined optimal density. After cells were attached, drugs were added using the Tecan D300e digital dispenser. A total of 10 μmol/L phenylarsine oxide (PAO) was used as a positive control and DMSO as a negative control. Cell viability assays were performed using CellTiter-Glow (Promega) according to the manufacturer's instructions and measured in the Envision (Perkin Elmer). Growth assays were plated as multiple measurements in the 384 plates (at least quadruple) and performed several times (at least three times). Measurements were normalized to the positive and negative controls. Combination index (CI) synergy scores were calculated as described (17 (link)), and displayed are the median CI scores from the synergy matrix (5 × 5 concentration range matrix, with twofold dilution steps). CI scores are defined as <0.1 very strong synergism; 0.1 to 0.3 strong synergism; 0.3 to 0.7 synergism; 0.7 to 0.85 moderate synergism; and 0.85 to 0.9 slight synergism.
9
High-throughput drug screening in glioblastoma
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We used a collection of 9 stem cell models of glioblastoma (HSR-GBM1 [23 (link)]; JHH520 [24 (link)]; NCH421k, NCH644 [25 (link),26 (link)]; BTSC-23, BTSC-233, BTSC-268, BTSC-349, BTSC-407 [27 (link)]), and exposed them to a library of clinical drugs (231 substances, each in 9 different concentrations; 4.33 nM to 25 M) using a semi-automated screening platform. For each cell model, repetitive drug resistance tests were performed: two biological (except for BTSC-23) and three technical replications. For each biological replication, the mean of the technical replications was used. Undifferentiated and immortalized human neural progenitor cells (ReNcell®CX, Sigma-Aldrich, St Louis, MO, USA) and neural stem cells (H9-Derived, Gibco) were used as healthy controls to evaluate the toxicity of the drugs. Additionally, we also tested the inhibition efficiency using three different normal adult human dermal fibroblasts (NHDF-Ad, Lonza, Basel, Switzerland). Effects on cell growth were assessed 72 h after substance exposure using the CellTiterGlow® assay (Promega, Madison, WI, USA). All procedures were in consent with the local ethical commission oversights.
10
Cytotoxicity Assays for CDKN1B-Deficient Breast Cancer Cells
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T47D-CDKN1BKO and MDA-MB-415-CDKN1BKO cells were seeded on 12-well plates for mafosfamide and doxorubicin cytotoxicity assays at densities of 2000 cells/well (T47D) or 2500 cells/well (MDA-MB-415). One day after seeding, ranges of concentrations of mafosfamide (Niomech, 0–100 μM) or doxorubicin (Sigma, 0–33 μM) were added to the cells in triplicate. After 3 days of drug treatment, cell viability was measured with CellTiter-Blue (Promega) using a 96-well plate reader (Tecan). Cells were then washed with PBS, fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. Plates were imaged and cell densities were measured using Image J Colony Area plugin.
For shCCND1-ZR-75-1 cells and T47D cells overexpressing CCND1, cells were seeded at a density of 500 cells/well in a 384-well plate. 24 h after seeding, cells were treated with ranging concentrations (0–10 μM) of doxorubicin (MedChemExpress: HY-15142) for a period of 7 days and cell viability was measured using Cell-Titer Glow (Promega).
For shCCND1-ZR-75-1 cells and T47D cells overexpressing CCND1, cells were seeded at a density of 500 cells/well in a 384-well plate. 24 h after seeding, cells were treated with ranging concentrations (0–10 μM) of doxorubicin (MedChemExpress: HY-15142) for a period of 7 days and cell viability was measured using Cell-Titer Glow (Promega).
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