PBMCs were thawed in a 37 °C water bath and washed with 10 ml of complete (c)RPMI containing RPMI 1640 (Gibco, ThermoFisher, 11875093), 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco, ThermoFisher, 15140122), 1 mM sodium pyruvate (Gibco, ThermoFisher, 11360070), 2 mM l -glutamine (Gibco, ThermoFisher, 35050061), 1× nonessential amino acids (Gibco, ThermoFisher, 11140050), 15 mM HEPES (Gibco, ThermoFisher, 15630080). T cells were isolated from PBMCs by three sequential rounds of magnetic separation using CD3 Microbeads (Miltenyi Biotec, 130-050-101), according to the manufacturer’s instructions. PBMC and T cells were seeded at 1 × 106 PBMC in a 96-well flat-bottom plate and rested in cRPMI for 1 h. After resting, cells were stimulated for 24 h with Gibco Dynabeads Human T-Activator CD3/CD28 (ThermoFisher, 11131D) in a bead-to-cell ratio of 0.5:1 in simultaneous presence or absence of 2 mM 2-Fluoro-2-deoxy-d -glucose (2-FDG) (Sigma, F5006) at 37 °C, 5% CO2.
2 fluoro 2 deoxy d glucose 2 fdg
Manufactured by Merck Group
2-Fluoro-2-deoxy-d-glucose (2-FDG) is a synthetic glucose analog. It is a radioactive tracer molecule used in medical imaging procedures, particularly positron emission tomography (PET) scans.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using 2 fluoro 2 deoxy d glucose 2 fdg
1
T-cell Activation with 2-FDG
2
Activation of T Cells with 2-FDG
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PBMCs were thawed in a 37 °C water bath and washed with 10 ml of complete (c)RPMI containing RPMI 1640 (Gibco TM , ThermoFisher, 11875093), 10 % FBS, 100 U/ml penicillin and 100 µg/ml streptomycin (Gibco TM , ThermoFisher, 15140122), 1 mM sodium pyruvate (Gibco TM , ThermoFisher, 11360070), 2 mM L-glutamine (Gibco TM , ThermoFisher, 35050061), 1X nonessential amino acids (Gibco TM , ThermoFisher, 11140050), 15 mM HEPES (Gibco TM , ThermoFisher, 15630080). T cells were isolated from PBMCs by three sequential rounds of magnetic separation using CD3 Microbeads (Miltenyi Biotec, 130-050-101), according to the manufacturer's instructions. PBMC and T cells were seeded at 1x10 6 PBMC in a 96-well flat-bottom plate and rested in cRPMI for 1 hour. After resting, cells were stimulated for 24 h with Gibco™ Dynabeads™ Human T-Activator CD3/CD28 (ThermoFisher, 11131D) in a bead-to-cell ratio of 0.5:1 in simultaneous presence or absence of 2 mM 2-Fluoro-2-deoxy-D-glucose (2-FDG) (Sigma, F5006) at 37°C, 5% CO2.
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