Medium oxygen levels were measured in an SDR SensorDish Reader (PreSens). Cells were washed twice with PBS and changed to RPMI without glucose and glutamine (Biological Industries) and with 10% dialyzed FBS (Gibco) at 106 cells/mL. They were plated at 0.8 mL of culture/well on pre-warmed 24-well OxoDish (PreSens) plates with integrated oxygen sensors. Medium oxygen levels were measured for 40 minutes to control for inter-well variability. The measurements were then paused to add either vehicle or 0.5 μM PU-H71 to the corresponding wells, and the oxygen of the medium was monitored for 1 more hour, at which point the measurement was paused again to add either PBS or 2 mM glutamine (Gibco) to the corresponding wells. Measurements went on for up to a total of 4 hours under regular incubation conditions (37 °C, 5% CO2). The measurements after addition of substrate were normalized to 1 by dividing by the levels of each well before addition of substrate.
Glucose and glutamine
Manufactured by Sartorius
Glucose and glutamine are essential nutrients used to support the growth and metabolism of cells in cell culture systems. They serve as primary energy sources and building blocks for various cellular processes. This lab equipment product provides a reliable and consistent supply of these critical components to maintain optimal cell culture conditions.
Automatically generated - may contain errors
2 protocols using glucose and glutamine
1
Oxygen Dynamics in Cell Metabolism
2
Measuring Oxygen Dynamics in Cell Cultures
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Medium oxygen levels were measured in an SDR SensorDish Reader (PreSens). Cells were washed twice with PBS and changed to RPMI without glucose and glutamine (Biological Industries) and with 10% dialyzed FBS (Gibco) at 106 cells/mL. They were plated at 0.8 mL of culture/well on prewarmed 24-well OxoDish (PreSens) plates with integrated oxygen sensors. Medium oxygen levels were measured for 40 minutes to control for interwell variability. The measurements were then paused to add either vehicle or 0.5 μmol/L PU-H71 to the corresponding wells, and the oxygen of the medium was monitored for 1 more hour, at which point, the measurement was paused again to add either PBS or 2 mmol/L glutamine (Gibco) to the corresponding wells. Measurements went on for up to a total of 4 hours under regular incubation conditions (37°C, 5% CO2). The measurements after addition of substrate were normalized to 1 by dividing by the levels of each well before addition of substrate.
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