This chapter covers the chemistry and experimental approach for studying tethered bilayernext term lipid previous termmembranesnext term (tBLM) for incorporating the previous termsmall peptide transmembrane ion channelnext term alamethicin. It describes the chemical components developed by AMBRI and methods for optimizing the tBLM containing alamethicin in order to construct stable and sensitive previous termsensors. The sensornext term containing alamethicin previous termchannelsnext term was then probed with amiloride and its analogues in the μM concentration range. The potency of amiloride and analogues as previous termchannelnext term 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 previous termchannelnext term proteins, previous termsmallnext term molecule sensing and biodiscovery.