A magnetically controlled electrochemical sensing membrane for detection of insulin was fabricated by magnetic field directed self-assembly of electromagnetic molecularly imprinted polymers (EMMIPs). EMMIPs with excellent electrochemical performance and multiple recognition sites to insulin were synthesized by surface polymerization on the ternary Fe3O4@rGO/PANI nanoparticles (MGP NPs). The insulin-EMMIPs obtained by recognizing insulin from sample were assembled onto a magnetic glassy carbon electrode (MGCE) surface in order to form the magnetically controlled membrane under the magnetic field induction, and the membrane was peeled off from the electrode by removal of the magnet after electrochemical measurement. In this work, the biomimetic sensing membrane could be controlled more effectively and the electrode could be regenerated more conveniently. The novel magnetically controlled sensing membrane showed good selectivity and reproducibility for the determination of insulin with the detection limit reaching 17 pM (S/N = 3), and this limit of detection was further reduced to 3 pM by using the Fe(CN)(6) (3 /4) redox probe. It has potential applications in the fields of clinical diagnosis with real-time detection capability, high efficiency, and easy operation properties. (C) 2016 Elsevier Ltd. All rights reserved.