Herein the electrochemiluminescent behavior of a promising and cheap electrochemiluminescence (ECL) luminophore, Eosin yellowish (2',4',5',7'-tetrabromofluorescein, EYH2), was investigated. A stable and strong ECL emission of Eosin yellowish could be achieved at electrochemical pretreated glassy carbon electrode in mild acidic media. Based on proposed stable cathodic ECL of Eosin yellowish on the electrochemically pretreated GCE, some important bioactive molecules, such as H 2 O 2 , guanine, adenine, nicotinamide adenine dinucleotide (NADH), uric acid and vitamin C, were found as enhanced coreactants towards the cathodic ECL of Eosin yellowish.