Cells were seeded into 96 well white opaque plates (Greiner) at 2000 cells per well and incubated at 37°C and 5% CO2 overnight. Cells were treated with selected drugs at different final concentrations and incubated for another 72 hr except for the initial studies of Aurora Kinase inhibitors in FLX1, where incubation was 120 hr. After incubation, plates and CellTiter-Glo (CTG, Promega) reagent were allowed to equilibrate at room temperature on the bench for 30 min. The CTG assay was performed following the manufacturer’s instructions and measured with a SpectraMax i3 Multi-Mode Platform (Molecular Devices). All experiments were done in at least biological duplicate with three technical replicates per condition. When multiple individual siRNA were used, the results are shown averaged in a standard dose response curve. In addition, AUC is calculated for each siRNA using GraphPad Prizm and shown as a separate point.
Celltiter glo ctg
Manufactured by Promega
Sourced in United States
CellTiter-Glo is a quantitative assay that measures the amount of ATP present in metabolically active cells. It provides a luminescent readout that is proportional to the amount of ATP, which is directly correlated with the number of viable cells in culture.
Automatically generated - may contain errors
Lab products found in correlation
96 well white opaque plate, by Greiner (1 mentions)
Spectramax i3 multi mode platform, by Molecular Devices (1 mentions)
Countess 2 cell counter, by Thermo Fisher Scientific (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Ckx41 fluorescence microscope, by Olympus (1 mentions)
Cell culture flask, by Corning (1 mentions)
Cell culture plate, by Corning (1 mentions)
C11995500cp, by Thermo Fisher Scientific (1 mentions)
Raw264.7 cell, by American Type Culture Collection (1 mentions)
Ensight plate reader, by PerkinElmer (1 mentions)
Echo 550, by Beckman Coulter (1 mentions)
3h thymidine, by PerkinElmer (1 mentions)
Synergy h1, by Agilent Technologies (1 mentions)
Wst 1, by Roche (1 mentions)
Caspase glo assay, by Promega (1 mentions)
Annexin 5 pe, by BioLegend (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by BD (1 mentions)
Infinite 200 spectrophotometer, by Tecan (1 mentions)
Erlotinib, by Cayman Chemical (1 mentions)
21 protocols using celltiter glo ctg
1
Cell Viability Assay Using CellTiter-Glo
2
Dose-Dependent Cytotoxicity Assay for AML Cells
3
Quantifying Cell Viability and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Cat.#M5655, Sigma-Aldrich) was used as previously described (27 (link),28 (link)). Experiments involving NAC, samples were washed and re-suspended with PBS before adding to MTT. Celltiter-Glo (CTG) (Cat.#PR-G7573, Promega) was used according to the protocol provided by manufacturer. For Annexin-V staining, stroma NK.Tert cells (5 × 103/100 μl) (RCB2350, Riken Cell Bank, Japan) were seeded on a 96-well plate overnight followed by cryopreserved CLL patient-derived primary cells (5 × 104/100 μl), total volume 200 μl. Stroma NK.Tert cells were used to maintain CLL patient-derived primary cell survival. Following drug treatment, cells were stained with 10 μl of 10× Annexin-V binding buffer (100 mM HEPES, 40 mM KCl, 1.4 M NaCl, 7.5 mM MgCl2, 25 mM CaCl2 pH 7.4) with Annexin-V and Hoechst-33342 for 15 min at RT, fixed with Annexin-V fix buffer (4% formaldehyde, 0.5% glutaraldehyde in 1× Annexin-V binding buffer) for 10 min and neutralized with N2 buffer (1.7 M Tris, 1.25 M glycine, pH9.1). Samples were gated and analyzed based on live population (Hoechst+ Annexin V−) with BD-Fortessa flow cytometer as described by Villalobos-Ortiz et al. (30 (link)). Invitrogen Countess® II cell counter (Cat.#AMQAX1000R) was used to determine number of live cells with Trypan Blue exclusion test (Cat.#T10282) following drug treatment.
4
Evaluating Co-Culture Media Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The co-culture media at 1:1 ratio of target cell (NPC43, HK1, C666–1, C17, B110, and G517) media and effector cell (NK-92) media was evaluated for any deleterious effect on the growth of target cells during co-culture. The target cells were first seeded into a 24- or 96-well plate in their own respective culture medium until cell attachment. Cell viability of the target cells in individual culture medium and in co-culture medium at 0 and 72 h was validated by using CellTiter-Glo (CTG; Promega, USA, #G7571), an established cell viability assay. Cell morphology was also observed by using Olympus CKX41 fluorescence microscope and bright field images were acquired at 24, 48 and 72 h.
5
RAW264.7 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW264.7 cells were purchased from the American Type Culture Collection. The culture reagents included Dulbecco's modified Eagle medium (DMEM; Gibco, C11995500CP), Roswell Park Memorial Institute (RPMI) 1640 (Gibco, C11875500BT), fetal bovine serum (Bio IND, 04-002-1A), antimycotic (Lifetechnologies, 15240-112), phosphate-buffered saline (PBS), pH 7.4 (Gibco, 10010-023), trypsin-EDTA (0.05%; Lifetechnologies, 25300-054), bovine serum albumin (Lifetechnologies, 15561012), and CellTiter-Glo (CTG; PROMEGA, G7572). Consumables and instruments included a cell culture plate (Corning), cell culture flask (Corning), microplate tester (BioTek, HM-1), and conventional instruments, such as a CO2 incubator (Thermo 3111) and biosafety cabinet (Heal Force, HFSAFe-1200LC).
6
High-throughput drug response screening in primary cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MNCs were added to pre-spotted drug plates (FIMM HTB)28 (link) or custom plates made with an Echo 550 (Beckman Coulter, Brea, CA, Supplementary Table 8 ). Cells were incubated in complete RPMI for 72 h (37 °C, 5% CO2). Cell viability was measured by CellTiterGlo (CTG, Promega, Madison, WI) on an EnSight plate reader (PerkinElmer, Waltham, MA). Drug sensitivity scores (DSS) were calculated with Breeze29 (link). Selective DSS (sDSS) values were calculated by subtracting healthy BM controls DSS values from patient DSS values. Fold growth was measured in untreated cells as the ratio of luminescence signal at 0 h and 72 h.
7
MM Cell Culture and Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human MM cell lines (HMMCL) were cultured in RPMI 1640 (Mediatech, Herndon, VA) supplemented with 10% fetal bovine serum (FBS). MM cell growth was measured by a 3(H)thymidine (PerkinElmer, Boston, MA) incorporation assay. Cell viability was analyzed by CellTiter-Glo (CTG) (Promega). Statistical significance was determined by Student’s t-test.
8
Evaluating Cell Viability Using CTG and WST-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was determined by two different assays; CellTiter‐glo (CTG) (Promega Corp., Madison, WI, USA) and cell proliferation reagent WST‐1 (Roche Diagnostics GmbH, Mannheim, Germany). CTG assay measures the cellular ATP levels as an indicator of metabolically active and viable cells. WST‐1 assay is dependent on NAD(P)H production by glycolysis and the activity of mitochondrial dehydrogenase enzymes, an indicator of metabolically active viable cells. Both the assays were performed as per the manufacturer's recommendations. CTG assay was performed in polystyrene 96‐well plates (Nunc, Thermo Fisher Scientific Inc., Paisley, UK) and luminescence was measured with Synergy H1 hybrid plate‐reader (BioTek, Winooski, VT, USA). The WST‐1 assay was performed in clear bottom 96‐well plates and the absorbance was measured at 450 nm.
9
Cell Viability, Apoptosis, and Caspase Assay
10
Compound Cytotoxicity Screening Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Compound cytotoxicity was assessed in a BSL-2 counter screen as follows using the Cell Titer-Glo Luminescent Cell Viability Assay (Severson et al., 2007 ). Host cells in media were added in 25μl aliquots (4000 cells/well) to each well of assay ready plates prepared with test compounds as above. Cells only (100% viability) and cells treated with hyamine at 100μM final concentration (0% viability) serve as the high and low signal controls, respectively, for cytotoxic effect in the assay. DMSO was maintained at a constant concentration for all wells (0.3%) as dictated by the dilution factor of stock test compound concentrations. After incubating plates at 37°C/5%CO2 and 90% humidity for 72 h, 30μl CellTiter Glo (CTG) (G7573, Promega) was added to each well. Luminescence was read using a BMG CLARIOstar plate reader following incubation at room temperature for 10 min to measure cell viability.
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!