The pharmacological properties of slow Ca²⁺-activated K⁺ current (K_slow) were investigated in mouse pancreatic β-cells and islets to understand how K_slow contributes to the control of islet bursting, [Ca²⁺]_i oscillations, and insulin secretion. K_slow was insensitive to apamin or the K_ATP channel inhibitor tolbutamide, but UCL 1684, a potent and selective nonpeptide SK channel blocker reduced the amplitude of K_slow tail current in voltage-clamped mouse β-cells. K_slow was also selectively and reversibly inhibited by the class III antiarrythmic agent azimilide (AZ). In isolated β-cells or islets, pharmacologic inhibition of K_slow by UCL 1684 or AZ depolarized β-cell silent phase potential, increased action potential firing, raised [Ca²⁺]_i, and enhanced glucose-dependent insulin secretion. AZ inhibition of K_slow also supported mediation by SK, rather than cardiac-like slow delayed rectifier channels since bath application of AZ to HEK 293 cells expressing SK3 cDNA reduced SK current. Further, AZ-sensitive K_slow current was extant in β-cells from KCNQ1 or KCNE1 null mice lacking cardiac slow delayed rectifier currents. These results strongly support a functional role for SK channel-mediated K_slow current in β-cells, and suggest that drugs that target SK channels may represent a new approach for increasing glucose-dependent insulin secretion. The apamin insensitivity of β-cell SK current suggests that β-cells express a unique SK splice variant or a novel heteromultimer consisting of different SK subunits.