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"Turn on" fluorescence immunoassay of endocrine diethyl phthalate. [Display omitted] • Fluorescence turn-on immunoassay of endocrine diethyl phthalate. • Red carbon dots can reduce interference of autofluorescence. • Red carbon dots as tags can improve the sensitivity of immunoassay. • "Turn-on" probe can reduce the interference of actual sample medium. Diethyl phthalate (DEP), as the environmental endocrine, would interfere with people's endocrine system and has greatly harm in health. Here a sensitivity, eco-friendly fluorescence immunoassay technique was established based on "turn-on" red carbon dots (R-CDs) conjugated anti-DEP antibody (Ab) as the immunoprobe for the detection of DEP in daily supplies. The R-CDs will "turn-on" fluorescence when it in formamide solution, due to the change of the electron distribution on the surface and electronic charge transfer of the red carbon dots. The use of "turn-on" R-CDs as immunoprobe effectively increased the sensitivity of the immunoassay. Moreover, the R-CDs conjugated anti-DEP Ab without any additional embellishment. Under the optimal conditions, a good linear was established with the DEP concentration range of 5 × 10−2 ng/mL–10 ng/mL, the low limit of detection (LOD) of the proposed method was 0.0056 ng/mL. This project has satisfactory accuracy with recoveries ranging from 96.0% to 115.1%. Our research provides an accurately, simply and sensitivity strategy to detect the content of environmental endocrine DEP in daily supplies. [ABSTRACT FROM AUTHOR]