CD2AP is targeted to the adherens junction along with E-cadherin after a calcium-switch assay. Incubation of MDCK cells without calcium for 3 h results in diffuse cytoplasmic localization of CD2AP that is distinct from E-cadherin localization. Upon replenishment of calcium, CD2AP and E-cadherin appeared at nascent cell–cell contacts within 30 min. By 1 h, most cells reformed cell–cell contacts with E-cadherin and CD2AP localization at the junction.