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Celery is acknowledged as a major food allergen in Europe, and mandatory labeling for preprocessed foods has been implemented. However, no methods for the specific detection of celery protein in foods have been published. In the present study, a sandwich celery ELISA using polyclonal anticelery antibodies for capture and detection was developed and validated. The method has an LOD of 0.5 mg/kg in buffer; however, it is applicable only for the screening of food products because of extensive cross-reactivity with potato and carrot proteins. Using nanoLC-ion-trap MS/MS, a number of proteins in the three vegetable species were identified as candidates for causing cross-reactions due to amino acid sequence homologies. Among others, a novel patatin (Sola t 1)-like protein was detected in celery and a flavin adenine dinucleotide binding domain-containing protein (Api g 5)-like protein was identified in carrot. The utility of triple-quadrupole MS/MS for specific and quantitative analysis of celery, potato, and carrot allergens was evaluated using whole protein extracts. Several unique precursor ion-to-product ion transitions were determined for each species, suggesting the feasibility of developing an MS-based screening method to specifically detect celery allergens in foods.