Ethics statement: All mice (C57Bl6/J, male and female) were purchased from Jackson Labs, housed and underwent drug treatment according to animal care and use protocol #4091, approved by the Temple University Institutional Animal Care and Use Committee, wherein all efforts were made to minimize suffering. The mice were given filter-sterilized i.p. injections of either vehicle (0.1% DMSO [D 4540, Sigma, St. Louis, MO] in PBS) or gefitinib (Gef, 5 mg/kg [G-4408, LC Laboratories, Woburn, MA]) followed 10 min later by an additional i.p. injection of either vehicle (sterile phosphate buffered saline [PBS]) or isoproterenol (ISO, 1 mg/kg [I6504, Sigma, St. Louis, MO]). After either 10 or 60 min the mice were euthanized according to the approved protocol above, their hearts were excised and their left ventricular tissue isolated, snap frozen in liquid N2 and stored at −80°C until further analysis.
G 4408
Manufactured by LC Laboratories
The G-4408 is a laboratory instrument designed for general analytical purposes. It features precision engineering and high-quality components to ensure reliable performance. The core function of the G-4408 is to facilitate various analytical procedures and measurements within a controlled laboratory environment.
Automatically generated - may contain errors
4 protocols using g 4408
1
Gefitinib and Isoproterenol Cardiac Tissue
2
Cytotoxicity Assay of Drug Compounds
3
High-Throughput Cell Viability Screening
4
High-Throughput Cell Viability Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Briefly, cells were plated in 384-well plates in 8 μL of media. Test compounds and dabrafenib (pharmacological assay control) were prepared as 10-point, 1:3 serial dilutions starting at 300 μM, then added to the cells (4 μL per well) using the Biomek NXP. Plates were incubated for 72 h at 37 °C, 5% CO2, and 95% relative humidity. After incubation, 4 μL of CellTiter-Glo® (Promega cat#: G7570) were added to each well, and incubated for 15 min at room temperature. Luminescence was recorded using a Biotek Synergy H4 multimode microplate reader. Viability was expressed as a percentage relative to wells containing media only (0%) and wells containing cells treated with DMSO only (100%). Three parameters were calculated on a per-plate basis: (a) the signal-to-background ratio (S/B); (b) the coefficient for variation [CV; CV = (standard deviation/mean) × 100)] for all compound test wells; and (c) the Z’-factor (18). The IC50 value of the pharmacological control (dabrafenib, LC Laboratories # G-4408) was also calculated to ascertain the assay robustness.
In case of viability rescue assays, cells were pre-treated with caspase, calpain, and autophagy inhibitors for 1–3 h before addition of test compounds. Time course viability assay was done with luminescence measurements performed at 4, 24, 48, and 72 h.
In case of viability rescue assays, cells were pre-treated with caspase, calpain, and autophagy inhibitors for 1–3 h before addition of test compounds. Time course viability assay was done with luminescence measurements performed at 4, 24, 48, and 72 h.
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