We present an electrodeposition method for fabricating thiolated polymer-based hydrogels through oxidation of hydroquinone (HQ). HQ is oxidized to benzoquinone (BQ) at an electrode, and the generated BQ is utilized for disulfide formation through an electrocatalytic reaction. As a simple demonstration of bioapplications, an electrodeposited hydrogel containing glucose oxidase was applied for glucose sensing. To the best of our knowledge, this is the first report on the electrodeposition of thiolated polymer-based hydrogels via disulfide formation using the indirect approach.