The physicochemical properties of palm olein fractions obtained by dry fractionation were studied. Chromatographic purification was applied to separate diacylglycerol from palm olein fractions without changing the triacylglycerol composition, and the effects of diacylglycerol on the cold stability of palm olein fractions were investigated. The cold test results proved that the influence of diacylglycerol on the crystallization behavior of palm olein fractions depended both on its concentration and the temperature of the system. Finally, the influencing mechanisms of diacylglycerol on the cold stability of palm olein fractions were proposed and discussed. Essential information for food application purposes in the palm oil industry is provided.