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Celltiter glo assay

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The CellTiter-Glo assay is a luminescent cell viability assay that quantifies the amount of ATP present in metabolically active cells. It provides a homogeneous method to determine the number of viable cells in culture. The assay is based on the ability of the luciferase enzyme to catalyze the oxidation of luciferin, which generates a luminescent signal that is proportional to the amount of ATP present.

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Lab products found in correlation

Caspase glo 3 7 assay, by Promega (21 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (13 mentions) Sytox green, by Thermo Fisher Scientific (12 mentions) Prism 6, by GraphPad (10 mentions) Prism 5, by GraphPad (10 mentions) Prism 7, by GraphPad (9 mentions) Envision, by PerkinElmer (8 mentions) Envision plate reader, by PerkinElmer (8 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions) Opti mem, by Thermo Fisher Scientific (6 mentions) Prism software, by GraphPad (6 mentions) 96 well plate, by Greiner (5 mentions) Synergy h4 hybrid reader, by Agilent Technologies (5 mentions) Synergy h1 plate reader, by Agilent Technologies (5 mentions) Microplate reader, by Tecan (5 mentions) Calcusyn software, by Biosoft (4 mentions) Glomax multi detection system, by Promega (4 mentions) Glomax plate reader, by Promega (4 mentions) Glomax discover system, by Promega (4 mentions)

838 protocols using celltiter glo assay

1

Evaluating CB's Cytotoxicity in Breast Cancer

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Breast cancer cells were seeded in 96-well plates at a density of 5 × 103 cells/well and after 20–24 h of incubation, cells were treated either with DMSO alone (0.02%, vehicle control) or with varying concentrations of CB (0.5–20 µM) in DMSO for an additional 24, 48, and 72 h in a CO2 incubator at 37 °C. For Nano-CB cell viability, CB is mixed with 20% intralipid at desired concentrations. Cell viability was assessed by using CellTiter-Glo assays (Promega Inc.). For combination studies, MDA-MB-231 cells are treated with vehicle, low dose of CB (200 nM), DOX (5 nM), and CB + DOX combination for 72 h. Cell viability was assessed by using CellTiter-Glo assays (Promega Inc.).
Rajamanickam S., Park J.H., Subbarayalu P., Timilsina S., Bates K., Yadav P., Nirzhor S.S., Eedunuri V., Mohammad T.A., Jung K.H., Onyeagucha B., Abdelfattah N., Benevides R., Lee G., Chen Y., Vadlamudi R., Brenner A., Kaklamani V., Jatoi I., Kuhn J., Hromas R., Gupta Y.K., Kaipparettu B.A., Arbiser J.L, & Rao M.K. (2022). Targeting aberrant replication and DNA repair events for treating breast cancers. Communications Biology, 5, 493.
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2

Analyzing Mycobacterium tuberculosis-Induced Cell Death

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TUNEL staining: Transfected MDMs on coverslips were infected with mCherry M.tb at MOI 5 and 50 for 24 and 48 h. Cells were then fixed with 4% PFA (Affymetrix, Santa Clara, CA) and labeled using the Click-iT TUNEL Alexa Fluor Imaging Assay (Invitrogen) following the manufacturer’s instructions. Cells were imaged with an Olympus FV1000 confocal microscope (Olympus, Shinjuku, Japan). Using Olympus Fluoview Viewer, at least 100 MDMs were manually counted to quantify % MDMs that stained with TUNEL.
CellTiter Glo Assays: Transfected or inhibitor treated MDMs in 96 well plates were infected with M.tb at MOI 5 for 24 h and cell death was assayed in triplicate with the CellTiter Glo Assay (Promega) following the manufacturer’s instructions.
Arnett E., Weaver A.M., Woodyard K.C., Montoya M.J., Li M., Hoang K.V., Hayhurst A., Azad A.K, & Schlesinger L.S. (2018). PPARγ is critical for Mycobacterium tuberculosis induction of Mcl-1 and limitation of human macrophage apoptosis. PLoS Pathogens, 14(6), e1007100.
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3

Compound 1 Selectivity for Myc-Expressing Cancer Cells

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Example 3

The selectivity of Compound 1 for cells expressing Myc over non-Myc expressing cells in cancer cell line P493-6 (engineered cell line derived from human B-cells) was determined. Adding doxycycline to P493-6 growth media shuts off Myc transcription [9]. Exemplary results are shown in FIGS. 3A to 3B, where cell viability was assessed by CELL TITER GLO assay (Promega). FIG. 3A depicts the assay results for cells expressing Myc, and FIG. 3B depicts the assay results for non-Myc expressing cells. Compound 1 demonstrated a selectivity for cells expressing Myc and exhibited decreased cell viability, compared to non-Myc expressing cells in cancer cell lines. The selectivity of other compounds described herein in cancer cell line cancer cell line P493-6 was also determined, and exemplary results are shown in Table 2, where viability was assessed by CELL TITER GLO assay (Promega).

US10865213B2. Max binders as MYC modulators and uses thereof (2020-12-15). Massachusetts Institute of Technology [US]. Inventors: Angela N. Koehler [US], Eric Stefan [US], Francisco Caballero [US], Dylan Vijith Neel [US], Nicholas B. Struntz [US], Helen L. Evans [US], Andrew Chen [US].
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4

Evaluating Compound 1's Effects on Cancer Cell Lines

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Example 2

The effect of Compound 1 on cell viability in several cancer cell lines and normal human astrocytes (NHA) was determined. The cancer cell lines employed in the assay included those corresponding to Burkitt's lymphoma (ST486 and CA46), small cell lung cancer (NCI-H1963), and glioblastoma (GBM4). The cell viability of normal human astrocytes (NHA) was also assayed. Exemplary results are shown in FIG. 2, where viability was assessed by CELL TITER GLO assay (Promega). Compound 1 demonstrated an effect in cell viability in a variety of cancer cell lines, consistent with the notion that Myc is a key oncoprotein in a broad range of cancers. The effects of other compounds described herein on cell viability in several cancer cell lines were also determined, and exemplary results are shown in Table 2, where viability was assessed by CELL TITER GLO assay (Promega).

US10106555B2. Max binders as MYC modulators and uses thereof (2018-10-23). Massachusetts Institute of Technology [US]. Inventors: Angela N. Koehler [US], Eric Stefan [US], Francisco Caballero [US], Dylan Vijith Neel [US], Nicholas B. Struntz [US], Helen L. Evans [US], Andrew Chen [US].
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5

Colony Assay and Cell Viability Measurement

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Cells were infected with lentiviruses encoding sgRNAs or treated with compounds as described. Colony assays were performed by replating cells at 500 cells per well in 6-well dishes and grown in regular growth media for 10 d before 100% methanol fixation, 0.05% crystal violet staining, and subsequent quantitation. CellTiter-Glo assay was performed as per the manufacturer’s instructions (Promega). Briefly, 1000 cells/well were plated into 96-well plates and treated with a range of compound concentrations. Cell viability was measured at the noted timepoints, based on luminescence by the CellTiter-Glo assay (Promega) and read on an Envision 2104 (PerkinElmer, USA), according to the manufacturer’s protocol.
Durbin A.D., Wang T., Wimalasena V.K., Zimmerman M.W., Li D., Dharia N.V., Mariani L., Shendy N.A., Nance S., Patel A.G., Shao Y., Mundada M., Maxham L., Park P.M., Sigua L.H., Morita K., Conway A.S., Robichaud A.L., Perez-Atayde A.R., Bikowitz M.J., Quinn T.R., Wiest O., Easton J., Schönbrunn E., Bulyk M.L., Abraham B.J., Stegmaier K., Look A.T, & Qi J. (2022). EP300 Selectively Controls the Enhancer Landscape of MYCN-Amplified Neuroblastoma. Cancer Discovery, 12(3), 730-751.
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6

Quantitative Assay of Cell Viability

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Cells were infected with lentiviruses encoding sgRNAs or treated with compounds as described. Colony assays were performed by replating cells at 500 cells per well in 6-well dishes and grown in regular growth media for 10 days before 100% methanol fixation, 0.05% crystal violet staining, and subsequent quantitation. The CellTiter-Glo assay was performed as per the manufacturer's instructions (Promega). Briefly, 1,000 cells per well were plated into 96-well plates and treated with a range of compound concentrations. Cell viability was measured at the noted time points based on luminescence by the CellTiter-Glo assay (Promega) and read on an Envision 2104 (PerkinElmer) according to the manufacturer's protocol.
Durbin A.D., Wang T., Wimalasena V.K., Zimmerman M.W., Li D., Dharia N.V., Mariani L., Shendy N.A., Nance S., Patel A.G., Shao Y., Mundada M., Maxham L., Park P.M., Sigua L.H., Morita K., Conway A.S., Robichaud A.L., Perez-Atayde A.R., Bikowitz M.J., Quinn T.R., Wiest O., Easton J., Schönbrunn E., Bulyk M.L., Abraham B.J., Stegmaier K., Look A.T, & Qi J. (2022). EP300 Selectively Controls the Enhancer Landscape of MYCN-Amplified Neuroblastoma. Cancer Discovery, 12(3), 730-751.
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7

Glioblastoma Stem Cell Viability Assay

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All viability assays were performed with GSCs below passage 20 using the CellTiter-Glo assay (Promega, WI, USA). Dose–response assays were performed in three step dilutions at six different concentrations in order to determine the growth inhibitory concentration required to inhibit 50% of cell proliferation (GI50). The 96-well plates were coated with matrigel (1:20, Trevigen, MD, USA) and seeded at 0.8-1.0×103 cells/well and were left to adhere for 24 hours. After 24 hours the compounds were added to the wells. Cell viability was measured on day five or eight after treatment by using the luminescent CellTiter-Glo assay (Promega, WI, USA) according to the manufacturer's protocol. Luminescence was measured with a Tecan Infinite Reader (Tecan Group Ltd., Männedorf, Switzerland). The GI50 values were calculated by median effect equation [34 (link)].
Venkatesan S., Hoogstraat M., Caljouw E., Pierson T., Spoor J.K., Zeneyedpour L., Dubbink H.J., Dekker L.J., van der Kaaij M., Kloezeman J., Berghauser Pont L.M., Besselink N.J., Luider T.M., Joore J., Martens J.W., Lamfers M.L., Sleijfer S, & Leenstra S. (2016). TP53 mutated glioblastoma stem-like cell cultures are sensitive to dual mTORC1/2 inhibition while resistance in TP53 wild type cultures can be overcome by combined inhibition of mTORC1/2 and Bcl-2. Oncotarget, 7(36), 58435-58444.
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8

Myc Selectivity Assay in P493-6 Cells

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Example 3

The selectivity of Compound 1 for cells expressing Myc over non-Myc expressing cells in cancer cell line P493-6 (engineered cell line derived from human B-cells) was determined. Adding doxycycline to P493-6 growth media shuts off Myc transcription [9]. Exemplary results are shown in FIGS. 3A to 3B, where cell viability was assessed by CELL TITER GLO assay (Promega). FIG. 3A depicts the assay results for cells expressing Myc, and FIG. 3B depicts the assay results for non-Myc expressing cells. Compound 1 demonstrated a selectivity for cells expressing Myc and exhibited decreased cell viability, compared to non-Myc expressing cells in cancer cell lines. The selectivity of other compounds described herein in cancer cell line cancer cell line P493-6 was also determined, and exemplary results are shown in Table 2, where viability was assessed by CELL TITER GLO assay (Promega).

US10106555B2. Max binders as MYC modulators and uses thereof (2018-10-23). Massachusetts Institute of Technology [US]. Inventors: Angela N. Koehler [US], Eric Stefan [US], Francisco Caballero [US], Dylan Vijith Neel [US], Nicholas B. Struntz [US], Helen L. Evans [US], Andrew Chen [US].
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9

Evaluating Compound 1 Effects on Cell Viability

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Example 2

The effect of Compound 1 on cell viability in several cancer cell lines and normal human astrocytes (NHA) was determined. The cancer cell lines employed in the assay included those corresponding to Burkitt's lymphoma (ST486 and CA46), small cell lung cancer (NCI-H1963), and glioblastoma (GBM4). The cell viability of normal human astrocytes (NHA) was also assayed. Exemplary results are shown in FIG. 2, where viability was assessed by CELL TITER GLO assay (Promega). Compound 1 demonstrated an effect in cell viability in a variety of cancer cell lines, consistent with the notion that Myc is a key oncoprotein in a broad range of cancers. The effects of other compounds described herein on cell viability in several cancer cell lines were also determined, and exemplary results are shown in Table 2, where viability was assessed by CELL TITER GLO assay (Promega).

US10865213B2. Max binders as MYC modulators and uses thereof (2020-12-15). Massachusetts Institute of Technology [US]. Inventors: Angela N. Koehler [US], Eric Stefan [US], Francisco Caballero [US], Dylan Vijith Neel [US], Nicholas B. Struntz [US], Helen L. Evans [US], Andrew Chen [US].
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10

Luminescence-based Cell Viability Assay

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Compound toxicity was measured by luminescence-based cell viability assay, CellTiter-Glo® Assay (Promega, Cat#G7570) was used according to the manufacturer’s instructions. Viability assays were performed in clear bottom 96-well plates (TPP, Cat#ZZ707902) where THP1-Cas9 cell cells were seeded at a cell density of 0.4 million cells/well and with the indicated doses of compounds. Cells were treated right after seeding. Luciferase measurement was performed after an indicated period (0–72 h) of compound incubation. To detect luciferase activity, CellTiter-Glo® Assay (Promega, Cat#G7570) was used according to the manufacturer’s protocol with minor changes. The reagent (50 µl) was added on top of the 50 µl cell-compound containing wells in a black with clear bottom 96-well plates (Greiner, Cat#655090) where the bottom of the plate was sealed (PerkinElmer, Cat#6005199). Luminescence was measured for 0.1 s with the EnVision® Multimode plate reader (Revvity, workstation version 1.14.3049.1193). Each of the experiments was repeated in three independent biological replicates. Fold increase in luminescence was calculated by dividing each read by its matching control group (DMSO treated), and results were plotted using GraphPad Prism 9.
Mutlu M., Schmidt I., Morrison A.I., Goretzki B., Freuler F., Begue D., Simic O., Pythoud N., Ahrne E., Kapps S., Roest S., Bonenfant D., Jeanpierre D., Tran T.T., Maher R., An S., Rietsch A., Nigsch F., Hofmann A., Reece-Hoyes J., Parker C.N, & Guerini D. (2024). Small molecule induced STING degradation facilitated by the HECT ligase HERC4. Nature Communications, 15, 4584.
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