TLR7/8 agonist 1 is a toll-like receptor ( TLR7 )/ TLR8 dual-agonistic imidazoquinoline. In Vitro TLR7/8 agonist 1 (Compound 5d) shows prominent immunostimulatory activities. TLR7/8 agonist 1 serves as a convenient precursor for the covalent attachment of fluorophores without significant loss of activity.TLR7/8 agonist 1 retains TLR7-agonistic activity with an EC 50 of 20 nM. TLR7/8 agonist 1 is covalently coupled directly to commercially-available fluorescein isothiocyanate and rhodamine B isothiocyanate. TLR7/8 agonist 1 (Compound 1) shows substantially different agonistic potencies in human TLR7 (50 nM) and TLR8 (55 nM) primary screens. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Cell Assay Fresh human peripheral blood mononuclear cells ( hPBMC ) are isolated from human blood obtained by venipuncture with informed consent and as per institutional guidelines on Ficoll–Hypaque gradients. Aliquots of PBMCs (10 5 cells in 100 μL/well) are stimulated for 12 h with graded concentrations of test compounds (e.g., TLR7/8 agonist 1; 0.1, 1, 10, and 100 μg/mL ). Supernatants are isolated by centrifugation and are assayed in duplicates using analyte-specific multiplexed cytokine/chemokine bead array assays. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Form:Solid IC50& Target:TLR7 TLR8