Ligand Activation: Mouse NFkB Secreted Alkaline Phosphatase SEAP - (SEAPorter?) Stable Reporter Cell Line [NBP2-26261] - TLR ligand stimulation assay. The RAW cell line was plated in 96-well plates at 5 x 10^4 cells/well. After 16 h, cells were stimulated with MALP-2, Pam3CSK4, Poly(I:C), LPS, Flagellin, R848 or mCpG as noted in each graph for 24 h. SEAP was analyzed using SEAPorter? Assay Kit.