Many hematophagous arthropods are stimulated by blood constituents to initiate feeding. We used a membrane-based feeding system to identify chemicals that stimulate acceptance and engorgement responses in various life stages of bed bugs. Water was fortified with a variety of compounds (e.g. salts, amino acids, vitamins, nucleotides, cholesterol and fatty acids) in these bioassays. ATP was the most effective phagostimulant in adults and nymphs, resulting in >70% of bed bugs fully engorging. Addition of NaCl to low ATP solutions that alone elicited <50% engorgement significantly enhanced feeding responses of bed bugs. A comparison of feeding responses with solutions of various adenine nucleotides showed that ATP was more stimulatory than ADP, which was more effective than AMP. Feeding assays with physiological levels of other blood constituents such as D-glucose, albumin, globulin, cholesterol and mixtures of vitamins and amino acids did not stimulate engorgement, suggesting that adenine nucleotides are the most important feeding stimulants in bed bugs. Identification of phagostimulants for bed bugs will contribute towards the development of artificial diets for rearing purposes, as well as for the development of alternative methods to eliminate bed bug infestations.