Luciferase Activity Assay

Lysate the cell using 1X Luciferase Cell Culture LysisReagent (Cat. E1531, Promega) following manufacturer instruction;

prepare Reaction Buffer (100µl for each sample): 500 µM luciferin (Cat E160E, Promega), 20 mM Tricine, 0.1 mM EDTA, 1.07 mM (MgCO3)4·Mg(OH)2, 2.67 mM MgSO4 in H2O, pH 7.8, with 33.3 mM DTT and 530 mM ATP.

put 20 µl/well of cell lysate into the white plate (Cat 3912, Corning);

turn on the luminometer (Veritas, Turner);

let the samples' plate and Reaction Buffer equilibrate at room temperature; Insert the plate into the instrument and prime the tube with the Reaction Buffer;

run the protocol on the luminometer (injection volume 100µl, delay before measurement 0s, integration time 10s)