The purpose of this study was to develop a protocol to increase freezing tolerance of field-grown ‘Chambourcin’ grapevines (Vitis spp.) using exogenous abscisic acid (ABA). The specific objectives were to determine the optimum concentration and timing for ABA foliar application in ‘Chambourcin’ and to evaluate morphological and physiological changes that lead to increased freezing tolerance in response to foliar ABA application. ‘Chambourcin’ grapevines were treated with a foliar ABA application of concentrations of 0, 100, 200, 300, 400, 500, 600, 700, and 800 mg·L−1 at 50% fruit set stage to evaluate ABA phytotoxicity under field conditions and identify the optimum concentration. In a subsequent experiment, ‘Chambourcin’ grapevines were treated with 400 and 600 mg·L−1 of ABA at different stages of development corresponding to 50% fruit set, 21 days after 50% fruit set, 50% veraison, 20, 30, 40, and 55 days postveraison. ABA concentrations of 700 and 800 mg·L−1 were phytotoxic and caused significant damage to leaves and flowers. Optimum concentrations of ABA did not affect yield components or basic fruit chemical composition, yet it promoted anthocyanin accumulation at harvest. Furthermore, ABA advanced bud dormancy, decreased bud water content, and eventually increased freezing tolerance under simulated freezing tests. The increased freezing tolerance of ABA-treated vines was confirmed by bud injury assessment after a natural freezing event in Jan. 2011. It was also determined that ABA was most effective when applied with an optimum concentration of 400 mg·L−1 20 to 30 days postveraison. It is concluded that exogenous ABA enhanced dormancy and increased freezing tolerance; thus, it has the potential to protect grape cultivars from freezing injury.