Revvity Alphalisa Surefire Ultra Phospho-Mlkl (Ser358)

This control lysate is intended for use with the AlphaLISA SureFire Ultra Phospho-MLKL (Ser358) Detection Kit or for use as a control for those running orthogonal or secondary assays along with the SureFire assay. To prepare the lysate, U937 cells were cultured in T175 flasks to 106 cells/mL in RPMI containing 10% FBS. Cells were treated with 20 μM Z-VAD for 30 minutes at 37°C, and then treated with 100 nM SM-164 and 20 ng/mL TNFα for 4 hours at 37°C. Total stimulus time 4.5 hours. Cells were then washed with HBSS and lysed in 5 mL 1X SureFire Ultra Lysis buffer.