Abstract Background Hydrogen peroxide (H 2 O 2 ) is a key reactive oxygen species (ROS) generated during appendage regeneration among vertebrates. However, its role during tail regeneration in axolotl as redox signaling molecule is unclear. Results Treatment with exogenous H 2 O 2 rescues inhibitory effects of apocynin‐induced growth suppression in tail blastema cells leading to cell proliferation. H 2 O 2 also promotes recruitment of immune cells, regulate the activation of AKT kinase and Agr2 expression during blastema formation. Additionally, ROS/H 2 O 2 regulates the expression and transcriptional activity of Yap1 and its target genes Ctgf and Areg . Conclusions These results show that H 2 O 2 is necessary and sufficient to promote tail regeneration in axolotls. Additionally, Akt signaling and Agr2 were identified as ROS targets, suggesting that ROS/H 2 O 2 is likely to regulate epimorphic regeneration through these signaling pathways. In addition, ROS/H 2 O 2 ‐dependent‐Yap1 activity is required during tail regeneration.