This chapter covers the chemistry and experimental approach for studying tethered bilayer lipid membranes (tBLM) for incorporating the small peptide transmembrane ion channel alamethicin. It describes the chemical components developed by AMBRI and methods for optimizing the tBLM containing alamethicin in order to construct stable and sensitive sensors. The sensor containing alamethicin channels was then probed with amiloride and its analogues in the μM concentration range. The potency of amiloride and analogues as channel blocker is rated against their molecular structure. Current research activities in related areas are also reviewed. The potential applications are studying "signature sequences" of large menbrane channel proteins, small molecule sensing and biodiscovery.