Calcium-binding proteins (CBPs) are ubiquitous pollen allergens and important food allergens in fish and amphibians. Calcium-binding allergens containing two EF-hands (polcalcins) have been detected and characterized in pollen from trees, grasses, and weeds. Timothy grass Phl p 7 is the most cross-reactive allergen among polcalcins. Although there is cross-reactivity described within the subfamilies of calcium-binding allergens, there are no strong indications for IgE cross-reactivity between CBPs from plants, fish, and humans. Therefore, Phl p 7 could be used as marker to identify multiple pollen-sensitized patients, whereas cod Gad c 1 or carp Cyp c 1 could be selected for the diagnosis of fish allergy. Hom s 4, a calcium-binding autoantigen, might be an interesting candidate to monitor chronic skin inflammation in atopic and nonatopic individuals. Diagnostic tests containing these molecules could allow the identification of most patients sensitized to calcium-binding allergens/antigens. In general, IgE recognition of calcium-binding allergens is influenced by binding or release of calcium ions. This knowledge could be used to engineer hypoallergenic CBPs for specific immunotherapy.