The present study was done to develop and evaluate a matrix transdermal patch for bisoprolol fumarate. Different combinations of Eudragit RS 100 and HPMC E5 were used with polyethylene glycol 400 as a plasticizer on a polyvinyl alcohol backing layer by the solvent evaporation technique. The patches were evaluated for organoleptic characteristics and physicochemical parameters. Initial in vitro dissolution experiments were conducted to optimize formulation parameters prior to ex vivo skin permeation studies. Eudragit RS 100 and HPMC E5 (9:1) combination was studied for skin permeation because of the sustain release effect. The effect of control patch and permeation enhancer including Tween 80, propylene glycol, and DMSO were evaluated at 10%-40% concentration in the Franz diffusion cell using excised abdominal skin of rabbit. Different kinetic models were used to interpret the release kinetics and drug release mechanism. The patch M04-PE containing 40% Tween 80 had better sustained release effect and had closer flux to the desired flux. M04-PE followed the zero-order kinetics with super case II release drug mechanism.