Cloning and expression of the recombinant canine NGAL protein used to raise the antibodies. (A) A schematic illustration of the NGAL construct is shown. The open reading frame of the canine NGAL was inserted into the vector pET32 downstream of the thioredoxin gene and linked to six histines. The size of recombinant canine NGAL protein was approximately 42 kDa. (B) The expression of canine NGAL was induced by IPTG in the host bacteria E. coli. As indicated by the arrowhead, recombinant NGAL, with a predicted size of approximate 42 kDa, was expressed (lane 2) and purified using Ni-NTA chromatography (lane 3). The identity of the recombinant NGAL protein (lane 2) was initially identified by Western blotting using an antibody against the His tag (C). M: protein size markers, lane 1: non-induction cell lysate.
Lee et al. BMC Veterinary Research 2012 8:248 doi:10.1186/1746-6148-8-248